A literature survey of studies performed by master ... · performed by master students at Département de Médecine Traditionelle (DMT) ... Avdeling for farmakognosi Kjemisk seksjon

Hovedfagsoppgave for graden cand. pharm.

A literature survey of studies

performed by master students at

Département de Médecine

Traditionelle (DMT) in Bamako,

Mali

Grete Hope

Avdeling for farmakognosi

Kjemisk seksjon

Farmasøytisk institutt

Universitetet i Oslo

2005

2

I Acknowledgements:

Kristian Børresen

My parents

Berit Smestad Paulsen

Drissa Diallo

Bente Rasch

Silje, Valeria, Dung, Osman, Man, Line and the employees on the 3rd

floor.

Students and employees in Mali

3

II Table of contents:

I Acknowledgments……………………………………………........ 2

II Table of contents………………………………………………….. 3

III Abbreviations……………………………………………………... 7

1. Introduction……………………………………………………….. 8

1.1 Traditional medicine……………………………………………… 8

1.2 The NUFU projects……………………………………………….. 8

1.3 Mali and traditional medicine ……………………………………8

2. Objectives and methods…………………………………………... 10

2.1 Objectives…………………………………………………………..10

2.2 Literature………………………………………………………….. 10

2.3 Field work……………….………………………………………… 10

2.3.1 Objectives……………….…………………………………………..10

2.3.2 Description………………..………………………………………... 11

2.4 Selected plants…………………………………………………….. 11

2.4.1 Methods for analysis……………………………………………….. 11

2.4.2 Excluded plants…………………………………………………….. 11

3. Results…………. …………………………………………………. 12

3.1 Plants………………………………………………………………. 12

3.1.1 Aizoaceae………………………………………………….. 12

3.1.1.1 Glinus oppostifolium……………………………………….. 12

3.1.1.2 Limeum pterocarpum……………………………………….14

3.1.2 Anacardiaceae…………………………………………….. 15

3.1.2.1 Lannea microcarpa………………………………………… 15

3.1.2.2 Lannea velutina…………………………………………….. 17

3.1.2.3 Mangifera indica…………………………………………… 25

3.1.2.4 Spondias mombin…………………………………………... 30

3.1.3 Annonaceae………………………………………………... 32 3.1.3.1 Annona senegalensis……………………………………….. 32

3.1.4 Araliaceae…………………………………………………. 35 3.1.4.1 Cussonia arborea…………………………………………... 35

3.1.5 Arecaceae………………………………………………….. 39 3.1.5.1 Borassus flabellifer………………………………………… 39

3.1.6 Asclepiadaceae……………………………………………..40

3.1.6.1 Calotropis procera…………………………………………. 40

3.1.6.2 Leptadenia hastata…………………………………………. 41

3.1.6.3 Leptadenia pyrotechnica…………………………………… 41

3.1.7 Asteraceae…………………………………………………. 42 3.1.7.1 Vernonia colorata………………………………………….. 42

3.1.7.2 Vernonia kotschyana……………………………………….. 45

3.1.8 Balanitaceae……………………………………………….. 47 3.1.8.1 Balanites aegyptiaca……………………………………….. 47

3.1.9 Bignoniaceae………………………………………………. 50

3.1.9.1 Stereospermum kunthianum………………………………... 50

3.1.10 Bombacaceae……………………………………………… 52

4

3.1.10.1 Bombax costatum ………………………………………….. 52

3.1.11 Caesalpiniaceae…………………………………………… 54 3.1.11.1 Afzelia africana…………………………………………….. 54

3.1.11.2 Burkea africana……………………………………………..54

3.1.11.3 Cassia nigricans…………………………………………….56

3.1.11.4 Cassia sieberiana…………………………………………... 58

3.1.11.5 Cordyla pinnata……………………………………………. 63

3.1.11.6 Daniellia oliveri……………………………………………. 64

3.1.11.7 Piliostigma thonningii……………………………………… 66

3.1.12 Capparidaceae…………………………………………….. 68

3.1.12.1 Boscia senegalensis………………………………………... 68

3.1.12.2 Maerua crassifolia…………………………………………. 70

3.1.13 Celastraceae……………………………………………….. 71 3.1.13.1 Maytenus senegalensis……………………………………... 71

3.1.14 Cochlospermaceae…………………………………………71 3.1.14.1 Cochlospermum tinctorium………………………………… 71

3.1.15 Combretaceae……………………………………………... 73 3.1.15.1 Combretum ghasalense…………………………………….. 73

3.1.15.2 Combretum glutinosum…………………………………….. 75

3.1.15.3 Combretum molle…………………………………………... 77

3.1.15.4 Pteleopsis suberosa………………………………………… 78

3.1.15.5 Terminalia avicennoides…………………………………… 80

3.1.15.6 Terminalia macroptera…………………………………….. 81

3.1.16 Convolvulaceae……………………………………………. 82 3.1.16.1 Ipomoea asarifola………………………………………….. 82

3.1.17 Ebenaceae…………………………………………………. 83 3.1.17.1 Diospyros abyssincia………………………………………. 83

3.1.18 Euphorbiaceae…………………………………………….. 84

3.1.18.1 Bridelia ferruginea………………………………………….84

3.1.18.2 Euphorbia sudanica………………………………………... 85

3.1.18.3 Ricinus communis………………………………………….. 85

3.1.18.4 Securinega virosa…………………………………………...86

3.1.19 Fabaceae……………………………………………………88

3.1.19.1 Afrormosia laxiflora……………………………………….. 88

3.1.19.2 Bauhinia thoningii…………………………………………. 91

3.1.19.3 Ostryoderris stuhlmannii……………………………………93

3.1.19.4 Prosopis africana…………………………………………... 95

3.1.19.5 Pterocarpus erinaceus……………………………………... 97

3.1.19.6 Pterocarpus lucens………………………………………….100

3.1.19.7 Stylosanthes erecta………………………………………….100

3.1.20 Flacourtiaceae…………………………………………….. 103

3.1.20.1 Flacourtia flabescens………………………………………. 103

3.1.21 Hypericaceae……………………………………………… 104 3.1.21.1 Psorospermum senegalense………………………………... 104

3.1.22 Liliaceae…………………………………………………… 106

3.1.22.1 Aloe buteneri……………………………………………….. 106

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3.1.23 Lycoperdeae………………………………………………..106

3.1.23.1 Podaxon aegyptiacus………………………………………. 106

3.1.24 Lythraceae………………………………………………… 108

3.1.24.1 Lawsonia inermis…………………………………………... 108

3.1.25 Meliaceae………………………………………………….. 110 3.1.25.1 Khaya senegalensis………………………………………… 110

3.1.25.2 Trichilia emetica…………………………………………… 111

3.1.26 Mimosaceae……………………………………………….. 114

3.1.26.1 Acacia ataxacantha………………………………………… 114

3.1.26.2 Acacia dudgeoni…………………………………………….115

3.1.26.3 Acacia senegal……………………………………………... 116

3.1.26.4 Acacia seyal………………………………………………... 117

3.1.26.5 Entada africana……………………………………………. 118

3.1.26.6 Parkia biglobosa…………………………………………… 121

3.1.27 Moraceae…………………………………………………... 121

3.1.27.1 Ficus iteophylla…………………………………………….. 121

3.1.28 Moringaceae………………………………………………. 122

3.1.28.1 Moringa oleifera…………………………………………… 122

3.1.29 Olacaceae………………………………………………….. 126

3.1.29.1 Ximenia americana………………………………………… 126

3.1.30 Opiliaceae…………………………………………………..131

3.1.30.1 Opilia celtidifolia…………………………………………... 131

3.1.31 Oxalidaceae………………………………………………... 136

3.1.31.1 Biophytum petersianum……………………………………. 136

3.1.32 Papaveraceae……………………………………………… 139

3.1.32.1 Argemone mexicana………………………………………... 139

3.1.33 Papilionaceae……………………………………………… 141

3.1.33.1 Swartzia madagascariensis………………………………… 141

3.1.34 Poaceae…………………………………………………….. 142

3.1.34.1 Zea mays…………………………………………………… 142

3.1.35 Polygalaceae………………………………………………..142

3.1.35.1 Securidaca longepedunculata…………………………….... 142

3.1.36 Rhamnaceae………………………………………………..149

3.1.36.1 Zizyphus mauritiana………………………………………...149

3.1.36.2 Zizyhpus mucronata………………………………………... 154

3.1.37 Rubiaceae………………………………………………….. 155

3.1.37.1 Canthium acutiflorum……………………………………… 155

3.1.37.2 Crossopteryx febrifuga……………………………………...157

3.1.37.3 Feretia apodanthera……………………………………….. 159

3.1.37.4 Mitracarpus scaber………………………………………… 161

3.1.37.5 Mitragyna inermis………………………………………….. 162

3.1.38 Salvadoraceae……………………………………………... 163 3.1.38.1 Salvadora persica………………………………………….. 163

3.1.39 Sapotaceae………………………………………………… 163

3.1.39.1 Butyrospermum parkii……………………………………... 163

3.1.40 Sterculiaceae………………………………………………. 164

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3.1.40.1 Cola cordifolia……………………………………………... 164

3.1.41 Ulmaceae…………………………………………………... 167

3.1.41.1 Celtis integrifolia…………………………………………... 167

3.1.42 Vitaceae……………………………………………………. 168

3.1.42.1 Cissus quadrangualris……………………………………... 168

3.2 Table of phytochemical results (with explanations)……………. 171

4. Discussion and conclusions………………………………………. 179

5. References…………………………………………………………. 182

A. Appendix…………………………………………………………... 186

A.1 Phytochemical methods and reagents…………………………… 186

A.1.1 Methods……………………………………………………. 186

A.1.1.1 Alkaloids…………………………………………………… 186

A.1.1.2 Polyphenolic substances…………………………………… 187

A.1.1.3 Flavonoides………………………………………………… 187

A.1.1.4 Tannins……………………………………………………... 188

A.1.1.5 Saponines…………………………………………………... 188

A.1.1.6 Cardiac glycosides…………………………………………. 189

A.1.1.7 Sterols and/or triterpenes…………………………………... 189

A.1.1.8 Coumarins………………………………………………….. 189

A.1.1.9 Anthraquinones and derivates……………………………… 190

A.1.1.10 Cyanogenic glycosides……………………………………...190

A.1.1.11 Reducing compounds………………………………………. 191

A.1.1.12 Mono- and polysaccharides………………………………... 191

A.1.1.13 Mucilages…………………………………………………... 191

A.1.1.14 Tetrahydrocannabinols……………………………………... 191

A.1.1.15 Water extractible substances……………………………….. 191

A.1.1.16 Water content………………………………………………. 191

A.1.1.17 Tests involving ashes………………………………………. 192

A.1.1.18 Thin layer chromatography………………………………… 193

A.1.1.19 Antioxidant activity………………………………………... 193

A.1.2 Reagents…………………………………………………… 193

A.2 List of plants and authors………………………………………... 195

7

III Abbreviations

AI - Atherogen Index = (Total cholesterol/HDL)

Ara - Arabinose

ASA - Acetyl salicylic acid

CNRST – The National Centre for Scientific Technological Research

CNS - Central Nervous System

DCM - Dichloromethane

DMT – Departement de Médecine Traditionelle

DPPH – 1,1-diphenyl-2-picryl-hydrazyle

EtOAc - Ethyl acetate

EtOH - Ethanol

FID - Flame Ionization Detector (used for gas chromatography)

Fuc - Fucose

Gal - Galactose

GalA - Galacturonic acid

Glc - Glucose

GlcA - Glucuronic acid

H2 - Hydrogen gas

H2O – water

HDL - High Density Lipoprotein (cholesterol)

HPLC – High Pressure Liquid Chromatography

IC50 - Inhibition concentration of 50 % cells

ICH50 - Inhibition concentration of 50 % heamolysis

LD50 - Lethal Dose for 50 % death

Man - Mannose

mAU - milli-absortion units (used in HPLC chromatography)

NMR – Nuclear Magnetic Resonance

NUFU – The Norwegian Council for Higher Education’s Program for Development

Research and Education

O2 - Oxygen gas

Rha - Rhamnose

Rf - Retention factor

SGOT – serum Glutamate Pyruvate Transaminase

SGPT - serum glutamic pyruvic transaminase

sp. - species (one)

spp. - species (several)

UV – Ultra violet

WHO – World Health Organisation

Xyl - Xylose

8

1. INTRODUCTION

1.1 Traditional medicine

Traditional medicine refers to health practice, approaches, knowledge and beliefs

incorporating plant, animal and mineral based medicines to treat, diagnose and prevent

illnesses or maintain well-being. 80 % of the population in Africa uses traditional

medicine for primary health care. In Ghana, Mali, Nigeria and Zambia, the first line

treatment for 60 % of children with high fever resulting from malaria is the use of herbal

medicines at home. WHO launched a traditional medicine strategy in 2002. This strategy

is meant to help countries to document traditional medicines and remedies, and to ensure

the safety and efficacy of these remedies (WHO, 2003).

Cooperation between developing countries and western countries can be beneficial in

helping to fulfill the goals of the traditional medicine strategy. The University of Oslo has

been cooperating with The National Centre for Scientific Technological Research

(CNRST) in Mali since 1989 and more specifically with the Department of Traditional

Medicine (referred to as DMT in the rest of this thesis) since 1996. These projects have

been possible thanks to The Norwegian Council for Higher Education’s Program for

Development Research and Education (NUFU) (SIU, 2002).

1.2 The NUFU projects

Several NUFU projects involving traditional medicine have been completed or are still

going on in Mali. 1996-2001: Contribution to phytochemical and pharmacological studies

of six medicinal plants from the Gourma. 2002-2006: Medicinal plants in Mali:

Ethnobothany, phytochemistry and biological activity (SIP, 2001). During these periods

several Norwegian master students and professors have been to Mali performing both

field work and laboratory work. The leader of DMT, Drissa Diallo, has spent time in

Oslo, Norway working on his doctorate thesis and doing laboratory and administrative

work for the projects, while three doctorate students from Mali have spent periods

working on their thesis, taking classes and doing laboratory work here.

1.3 Mali and traditional medicine

Mali, the largest country in West Africa, is bordered by 7 countries. Algeria is situated to

the north and north east, Nigeria to the east, Burkina Faso to the south east. The Ivory

Coast is to the south, while Senegal and Mauritania are to the west. The capital is called

Bamako. Mali has no coast line, but the river Niger passes through most of the country so

only the northern areas are very dry. The northern desert parts are a part of the Sahara

desert. The number of inhabitants is nearly 12 million (2004) belonging to different

ethnical groups. Life expectancy at birth is 45,28 years. An average of 64 % of the

population is living below the poverty line (2001) (CIA, 2004). The most common

diseases are malaria and other infections, including schistosomiasis and fungal infections,

diabetes in some areas, and different kinds of wounds (SIU, 2002).

Mali was a French colony until 1960 and the use of traditional medicine was illegal under

9

French rule. Luckily the traditions were still conserved in secret so at least parts of the

knowledge still exists, and it is now being recorded by the DMT.

DMT has also developed several so called Improved Traditional Medicines (IMT). They

contain standardized amounts of one or more herbs with long tradition in traditional

medicine, and are a cheap alternative to pharmaceuticals from western countries. Current

IMTs are as follows:

Table 1.3.a Current IMTs used in Mali

Name Indication Plant(s)

Balembo Cough Crossopteryx febrifuga

Dysenteral Dysentery Euphorbia hirta

Psorospermin Dermatosis Psorospermum guineense

Hepatisan Hepatitis Combretum micranthum

Laxia-cassia Obstipation Cassia italica

Malarial Malaria Cassia occidentalis, Lippia

chevalierii, Spilanthes

oleracea

Gastrosedal Stomach ulcer Vernonia kotschyana

10

2. OBJECTIVES AND METHODS

2.1 Objectives

The main objective is to help improve the health care situation in Mali. This can for

example be done by checking plants for active and/or toxic ingredients, and by helping

with development of new ITMs.

More specific objectives of this thesis are:

- to compile and translate the results of some of the scientific work performed in Mali

between 1993 and 2003 by master students.

- to detect plants with medical properties that have not yet been thoroughly examined.

This thesis is meant to be a summary of the results from the examined theses. Because of

this a brief introduction of the respective theses is included for each plant.

2.2 Literature

15 master theses written from 1993 to 2003 at Department of Traditional Medicine

(DMT) in Bamako, Mali were examined. All theses were written in French so translation

errors might exist. Other sources were different articles and other literature survey theses

on some of the plants have been used (see results and reference list).

A short introduction of the known traditional uses of each plant is added if available.

Most of this information is found in the “Pharmacopée sénégalaise traditionelle – plantes

médciniales et toxiques” by Kerharo and Adam (1974). Senegal is Malis neighbour to the

west and the vegetation in the two countries is very similar. Small differences between

species might of course occur.

2.3 Field work

The project (see 1.2) comprises both field work and laboratory work. So even though this

thesis basically is a literature survey (theoretical), the stay in Mali included a field trip for

5 days.

2.3.1 Objectives

- Identify plants used against dermatosis.

- Identify the uses of 6 specific plants: - Biophytum petersianum (see 3.1.31.1)

- Cola cordifolia (see 3.1.40.1)

- Combretum glutinosum (see 3.1.15.2)

- Lannea velutina (see 3.1.2.2)

- Opilia celtidifolia (see 3.1.30.1)

- Ximenia americana (see 3.1.29.1)

11

2.3.2 Description The field work was done in the Kolokani district, 30 kilometers north of Bamako.

The following villages were visited: N’Tjobougou, Massantola and Didieni. Healers from

the villages and areas surrounding them were interviewed. They were first asked which

plants they used to treat dermatosis1. Then they were asked how they used the plants

mentioned in 2.3.1. All results are included in 3.1 under the respective plant.

N’Tjobougou: 5 healers were interviewed.

Didieni: 6 healers were interviewed.

Massantola: 14 healers were interviewed.

All interviews were performed using the local language Bambara and translated to

English by Drissa Diallo. Information might have been lost or misunderstood during

translation.

Results: se 3.1

2.4 Selection of plants

The selected plants consist of all plants subjected to one or more tests in the theses and all

plants recorded from the interviews during the field trip. The number of plants tested in

one thesis was between 1 and 18. Some were reviewed by Aasberg (2001) and Tran

(2004) and for these, only the results from the Malian theses were recorded. Diallo (2000)

performed an etnopharmacological survey of medicinal plants in Mali and phytochemical

study of four of the identified plants. This is noted after the respective plants.

For the rest of the plants a quick search using the commercial search site

www.google.com and the scientific search site www.PubMed.com was performed to

check the amount of available literature. This gives an indication of the amount of

available literature. The quality of content on the identified sites was not checked, but we

may consider the articles found using PubMed.com to be of higher scientific importance.

The plants are listed alphabetically after family, then after plant name. A total of 82

plants from 42 families were included.

Genus and family were identified by using W3TROPICOS

(http://mobot.mobot.org/W3T/Search/vast.html).

2.4.1 Methods for analysis

Extraction(s) are described for each plant.

The phytochemical tests performed are as far as possible described in the appendix (A.1).

The other tests can be found in the respective theses, some are however briefly described

in this thesis.

2.4.2 Excluded plants: From the field work Vitex spp. was identified (no species name). A survey of all Vitex

species is beyond the scope of this thesis.

1 Dermatosis is here defined as all diseases affecting the skin.

12

3. RESULTS

3.1 Plants

3.1.1 Aizoaceae

3.1.1.1 Glinus oppostifolius (L.) Aug. DC.

Thesis: Bah (1998)

Some traditional uses:

In Mali, pulverized stems with leaves are put into food as a treatment for abdominal pain

and jaundice. It is used together with Echinochloa stagnina (Rets.) P. Beauv. to treat

malaria, dizziness and to stimulate appetite (Diallo et al, 1999). It has also been used for

wound healing, as anti-inflammatory and analgesic agent and against diarrhoea (Diallo,

2000). Debes (Debes 1998) identified the following uses of this plant after interviews

with healers in Gao and Gourma in Mali: malaria, head ache, weight loss, nausea, pain

(externally and in the joints), skin-diseases, constipation, diarrhoea, wounds, jaundice,

stomach ache (parasites) and high fever. Malaira is the most cited indication for this

plant.

Bah (1998):

This author studied the larvicidal effects of 10 different plant extracts on Anopheles

gambiae, one of the intermediate hosts in malaria. As mosquitoes are developing

resistance to an increasing number of chemical insecticides, it is important to search for

new and possibly less toxic agents from plants to try and control the disease.

Anopheles gambiae mosquitoes were collected in Mopti. For the tests, larvae were

elevated and collected in the laboratory, from full grown females collected at one specific

date. Larvae were fed with Whiskas ® cat food.

The plant material was collected in Diré in June 1996, and was dried and pulverized

before analysis. The whole plant was used.

Extractions:

250 g plant material

Dichloromethane (DCM) 1500 ml x 3

_______________________/ \

DCM extract \

Residue

/ Methanol (1500 ml x 3)

________________________/ \

Methanol extract \

Residue

Figure 3.1.1.1.a Extraction with solvents of different polarity

13

50 g plant material + water (500 ml x 24 hours x 3)

/

Water extract

Figure 3.1.1.2.b Maceration

All extracts were concentrated and lyophilized before use. They were diluted with

distilled water to the appropriate concentrations before the larvicidal test was performed.

Tests:

Extracts were tested on groups of 8 x 25 larvae. The extracts were tested at

concentrations from 0,015 mg/ml to 0,06 mg/ml (preliminary tests) and from 1 mg/ml to

50 mg/ml later. Dead or dying larvae were registered as they floated to the surface of the

test container.

Well known chemical insecticides (ex. permethrine) were tested tested on full grown

mosquitos. The results from these tests can not be compared to the results from the tests

performed on the larvae.

Results:

- from the preliminary tests:

The DCM extracts gave 1 % mortality for 0,015 mg/ml and 0,03 mg/ml and 4 %

mortality for 0,06 mg/ml. Only the methanol extract at 0,06 mg/ml showed activity, with

1 % mortality. None of the water extracts showed any effect.

-from the extended tests:

Table 3.1.1.1.a Methanol extract

Concentration

mg/ml

Number of larvae

tested

Number of deaths Mortality (%)

1 200 42 21

5 125 35 28

10 200 49 24,5

25 200 67 33,5

50 200 81 40,5

Negative control 500 5 1

Table 3.1.1.1.b DCM extract

Concentration

mg/ml

Number of larvae

tested

Number of deaths Mortality (%)

1 200 18 9

5 200 39 19,5

10 200 60 20

25 200 85 42,5

50 200 140 70

Negative control 500 5 1

14

The DCM extracts show the strongest activity.

This plant showed the strongest insecticide activity together with Limeum pterocarpum

(3.1.1.2). This might be caused by saponins, terpenes or alkaloids formerly identified in

the plant. From the 10 tested plants this was the only one that went through extended

tests.

Diallo (2000) did a phytochemical study and a review of this plant. He found the same

larvicidal effect as Bah as well as antioxidant, antifungal and molluscicidal effects and

effect on the complement system. Inngjerdingen (2000) studied the structure and activity

of the polysaccharids found in this plant.

3.1.1.2 Limeum pterocarpum (Gay.) Heimerl.

Thesis: Bah (1998)


In Mali, leaves and stem are used against malaria. The whole plant is crushed and boiled

in water until it changes color. The solution is drunk for 3 days to cure malaria (Diallo et

al, 1999). In the Gourma-district in Mali, this plant is used to treat heart diseases and

scorpion-bites in addition to the use against malaria (Diallo et al, 1992).

Bah (1998) This author studied the larvicidal effects of 10 different plant extracts on Anopheles







The plant material was collected in Douentza in September 1995, and was dried and

pulverized before analysis. Twigs and fruits from L. pterocarpum were analyzed.

Extractions:



_______________________/ \

DCM extract \

Residue


________________________/ \

Methanol extract \

Residue

Figure 3.1.1.2a Extraction with solvents of different polarity

15


/

Water extract

Figure 3.1.1.2b Maceration

All extracts were concentrated and lyophilized before use. They were diluted with

distilled water to the appropriate concentrations before the test was performed.

Test:


concentrations from 0,015 mg/ml to 0,06 mg/ml. Dead or dying larvae were registered as

they floated to the surface of the test container.

Results:

DCM extracts: 0,015 mg/ml and 0,03 mg/ml gave respectively 1 % and 2 % mortality

while 0,06 mg/mg gave 6 % mortality.

Methanol extracts: 0,03 mg/ml and 0,06 mg/ml gave respectively 2 % and 2,5 %

mortality while the extract at 0,015 mg/ml showed no effect.

Water extracts: No activity was seen.

Even though this plant showed the best larvicidal activity of the 10 tested plants it did not

go through extended tests.

Diallo et al (1992) did a phytochemical screening of this plant. The following substances

were identified: coumarins, alkaloids (0,30 %), sterols and/or triterpenes, mono- and

polysaccharides, mucilages and cardiac glycosides.

Diallo et al (1999) identified traditional uses of this plant doing an ethnobotanical survey

in the Gourma district in Mali.

3.1.2 Anacardiaceae

3.1.2.1 Lannea microcarpa Engl.

Thesis: Sanogo (1999)


In Senegal, this plant is often confounded with and used in the same ways as Lannea

velutina A. Rich (see 3.1.2.2). It is however considered to have weaker effect than

L. velutina. Both plants are used against diarrhoea, externally in baths for children with

rickets, and for adults with generalized pain without any apparent cause. (Kerharo and

Adam, 1974).The gum from the plant has traditionally been used to treat dysentery

(Sanogo 1999).

16

Sanogo (1999) This author did a survey of gums found from different plants in Mali and their

commercial and medicinal uses. The collection of plant material took place between

November 1997 and February 1998 in Bamako and Banankoro. This specific gum was

collected in December 1997 in Banankoro.

Tests:

Standard phythochemical tests (A.1.1) were performed at DMT.

Thin layer chromatography was performed with known monosaccharides as reference

substances, and with the following solvent system: Ethyl acetate: Methanol: Acetic acid:

Water (13:3:4:3)

Gas chromatography was used to confirm the content of monosaccharides.

All gums were tested for possible content of Escherichia coli and Salmonella spp.

They were also examined for content of starch and starch derivatives.

Results:

From the phytochemical tests the following substances were found:

Coumarins, tannins, mono- and polysaccharides, mucilages, sterols and/or triterpenes,

and cardiac glycosides. The content of water was between 7 and 7,95 %. Ashes insoluble

in HCl were 3,29 %, total ash content was 3,69 % and sulphur containing ash was 5,69 %

(Table 3.2).

Thin layer chromatography:

The tested extracts gave matching Rfs with galactose, galacturonic acid and glucuronic

acid (they all had the same Rf here) and arabinose. In addition to this there was detected

one spot that did not correspond with any of the references sugars.

Gas chromatography:

7 monosaccharides: Galactose (70,38 %), arabinose (14,23 %), galacturonic acid (7,85

%), glucuronic acid (5,33 %), mannose (1,11 %), rhamnose (0,66 %) and glucose (0,44

%). Only the 4 largest groups were detected with the thin layer chromatography.

The tests for both Escherichia coli and Salmonella spp were negative for 1 g of gum.

The test for starch and starch derivatives was also negative.

Field work:

While asking about the uses of Lannea velutina this information was given:

A parasite on Lannea microcarpa is used for preparation of a decoction, this is used

against headache. The vapour from the boiling decoction is inhaled.

For more information about this plant see Aasberg (2001) and Diallo et al (Diallo 2002).

17

3.1.2.2 Lannea velutina A. Rich.

Theses: Kanta (1999), Ouologuem (1999) and Samake (2000).


In Senegal, preparations from the stem and roots are used internally against diarrhoea,

externally in baths for children with rickets and for adults with generalized pain without

any apparent cause (Kerharo and Adam, 1974).

Kanta (1999)

This author identified 29 plants used in the treatment of infections by Candida albicans,

These were identified after interviews of healers and salesmen of medicinal plants and a

review of available literature in the period between July and October 1998, using the

search words vaginal discharge and vaginitis. 18 of these plants were collected the 19th

of November 1998 in the Tienfala forest 30 km from Bamako and 10th

December 1998 in

mount Manding in Siby, 60 km from Bamako.

Stem bark from Lannea velutina was analyzed. An extract was made with ethanol and

tested for activity against Candida albicans and Escherichia coli, but it did not have any

effect against these microorganisms.

Ouologuem (1999)

This author tested the larvicidal effects of different extracts of 11 Malian plants. Larvae

from Anopheles gambiae and Culex quinquefasciatus were used. 6 of the plants were

tested on both species, while 5 were only tested on C. quinquefasciatus.

Larvae from second and third generation Anopheles gambiae collected in Mopti were

used. Larvae from Culex quinquefasciatus were collected in N’Tomikorobougou and

Darsalam, in the Bamako district. Leaves from Lannea velutina were collected in

Tienfala. The leaves from this plant were chosen for further testing because of their good

larvicidal properties (see results).

Extraction:

Drug 350 g

/ Ethanol

___________________ / \

Extract with ethanol \

/ Residue

/ H2O

____/

Ethanol extract dissolved in H2O

/ \

/ DCM \

___/ \___

DCM extract Ethanol + H2O extract


18

Tests:


Column chromatography was performed on the DCM-extract.

For the first column, the following solvent system was used: N-hexane : Isopropanole

(14:1 – 10:1 – 4:1 – 2:1 – 1:1 – 1:2).

The fractions were tested on thin layer chromatography with the following solvent

system: CHCl3:MeOH (10:1).

Interesting fractions were tested on a second column with solvent systems of MeOH and

CHCl3 at different concentrations. These fractions were also tested for larvicidal activity.

All fractions were tested using thin layer chromatography. The plates were treated with

Godins reagent and examined at 250 and 366 nm.

HPLC was performed at the University of Lausanne

Specifications:

Injected volume: 10 µl

A system of acetonitrile and water at different proportions were used; from 25-100% for

30 minutes, 100 % for 10 minutes and 100-25% for 30 minutes.

All tested solutions were dissolved in methanol at a concentration of 1 mg/ml.

NMR was performed at the University of Lausanne.

Specifications: 1H-NMR spectra were done at a frequency of 499 85 MHz.

All tested solutions were dissolved in pyridine deuteride 5.

Tetramethylsilane was used as internal standard. The values for the chemical

displacement were expressed in ppm compared to the internal standard.

Tests on larvae from Anopheles gambiae and Culex quinquefasciatus were done on 5,5

mg extract dissolved in 110 µl of a suitable solvent; water for the water and ethanol

extracts and dimethylsulfoxide (DMSO) for the methanol, ether and DCM-extracts. The

tests were performed on 5x20 larvae. Larvae that had fallen to the bottom of the tubes or

were immobile were considered dead.

The DCM extracts from Lannea velutina leaves and Cussonia arborea roots, methanol

extract from Diospyros abyssinica leaves and ether extract from Cissus quadrangularis

twigs were tested at different concentrations. The extended tests were only performed on

Culex quinquefascitus larvae, as these are considere to be more resistant than larvae from

Anopheles gambiae.

Results:

From the phytochemical tests performed at DMT the following substances were found:

Coumarins, tannins, mono- and polysaccharides, mucilages, sterols and/or triterpenes,

leucoanthyocyans, reducing compounds, flavonoids and cardiac glycosides. The content

of water was between 6 and 6,10 %. Ashes insoluble in HCl were 8,70 %, total ash

content was 5,2 % and sulphur containing ash was 8,0 % (Table 3.2).

19

Larvicidal tests:

Table 3.1.2.2.a Culex quinquefasciatus - % deaths at 500 ppm

Plant extract

Time of

exposure

L.v.l.

Methanol

L.v.l.

DCM

L.v.b.

Methanol

L.v.b.

DCM

L.v.r. H2O L.v.r.

DCM

30 minutes 5 95 0 0 10 0

1 hour 15 100 0 5 10 0

24 hours 40 100 65 15 35 10

Table 3.1.2.2.b Anopheles gambiae - % deaths at 500 ppm

Plant extract

Time of

exposure

L.v.l.

Methanol

L.v.l.

DCM

L.v.b.

Methanol

L.v.b.

DCM

L.v.r. H2O L.v.r.

DCM

30 minutes 85 50 0 80 90 95

1 hour 90 70 5 80 90 95

24 hours 100 100 60 100 95 100

L.v.l. Lannea velutina leaves

L.v.b. Lannea velutina bark

L.v.r. Lannea velutina roots

Extended tests at different concentrations:

L.v.l. Lannea velutina leaves

C.a Cussonia arborea roots

D.a.l. Diospyros abysscinia leaves

C.q. Cissus quadrangularis twigs

Table 3.1.2.2.c Culex quinquefasciatus - % deaths at 250 ppm

Plant extract

Time of

exposure

L.v.l.

DCM

C.a.

DCM

D.a.l.

Methanol

C.q.

Ether

Distilled

water

30 minutes 5 0 35 30 0

1 hour 15 25 40 40 0

24 hours 100 95 100 85 0

Table 3.1.2.2.d Culex quinquefasciatus - % deaths at 125 ppm

Plant extract

Time of

exposure

L.v.l.

DCM

C.a.

DCM

D.a.l.

Methanol

C.q.

Ether

Distilled

water

30 minutes 0 0 15 0 0

1 hour 30 5 30 0 0

24 hours 85 30 80 5 0

20

Table 3.1.2.2.e Culex quinquefasciatus - % deaths at 62,5 ppm

Plant extract

Time of

exposure

L.v.l.

DCM

C.a.

DCM

D.a.l.

Methanol

C.q.

Ether

Distilled

water

30 minutes 0 0 0 0 0

1 hour 0 0 0 0 0

24 hours 60 0 25 0 0

Table 3.1.2.2.f Culex quinquefasciatus - % deaths at 31,25 ppm

Plant extract

Time of

exposure

L.v.l.

DCM

C.a.

DCM

D.a.l.

Methanol

C.q.

Ether

Distilled

water

30 minutes 0 0 0 0 0

1 hour 0 0 0 0 0

24 hours 50 0 0 0 0

Column chromatography:

Fractions 2, 7, 9 and 11 from the first column were tested separately on column 2.

Fraction 2 seems to have good larvicidal effect at all tested times, followed by fraction 4.

Fraction 11 shows best effect after 24 hours.

HPLC:

A top was observed at 1050 mAU after 28,50 minutes of eluation and another top after 12

minutes of eluations. Chlorophyll was observed at 254 and 366 nm.

3 visible tops were observed at 366 mn, they came after 16 mn, 19,20 mn and 21 mn of

eluation. Most compounds were eluated between 10 and 30 minutes.

Substance LV5 (13 minutes of eluation) was isolated, its UV spectrum showed maximum

absorption at 210, 225.

NMR:

Substance LV5: the proton spectrum is characterized by aliphatic chains between 0 and 3

ppm, a sugar at 4,5 ppm and an aromatic ring at 7 ppm. This substance (LV5) seems to be

a glycosylated substance with lateral chains and an aromatic ring of triterpene type.

The DCM extract of Lannea velutina leaves showed the best larvicidal effect of the tested

plants.

Samake (2000)

This author identifed 13 plants and one mushroom used in wound healing and studied

their monosaccharide contents and activities on the complement system. The plants were

identified after interviews with local healers. All plants were collected in the forest near

Siby in November 1998, dried and pulverized before analyzis.

21

Extraction:

Plant material (10-30g)

/ Ethanol (80º)

________________________/ \


Residue

/ H2O 50 º C

___________________________/ \

Extract with H2O at 50 º C \

Residue

/ H2O 100 º C

_____________________________/ \

Extract with H2O at 100 º C Residue

Figure 3.1.2.2.b

A dialysis was performed on all extracts, followed by lyophilization.

Tests:

Thin layer chromatography was performed with known monosaccharides as references.

All solutions were hydrolyzed before testing to separate polysaccharides into separate

monosaccharides. The following solvent system was used: Ethyl acetate – Acetic acid –

Water – Methanol (13:3:3:4). The plates were treated with a mixture of ansidine,

phthalate and methanol (1,23 : 1,66 : 100 ml) and examined using UV-light at 366 nm.

Activity on the complement systems was tested on sheep erythrocytes.

Results:

Thin layer chromatography and effect on the complement system:

Table 3.1.2.2.g Extracts tested with known sugars as references and ICH50

Tested

substance

Tested

part

Ara Rha Xyl Fuc Man Gal Glc GalA GlcA ICH50

(µg/ml)

L.v.

50ºC

Stem

bark

+ + + 0 + + + + + 40

L.v.

100ºC

Stem

bark

+ + + 0 0 + + + + 50

L.v. = Lannea velutina

22

Gas chromatography:

Table 3.1.2.2.h Amounts of monosaccharides and polysaccharides detected

Monosaccharides Extract at 50ºC Extract at 100ºC

Sugar ratio (%) Sugar ratio (%)

Arabinose 19 24

Rhamnose 12 8

Xylose 1 0

Mannose 2 2

Galactose 47 38

Glucose 2 15

Glucuronic acid 5 3

Galacturnoic

acid

12 10

Total 100 100

Polysaccharide

content

81 % 47 %

Field work:

N`Tjobougou, 5.2.04

Djokin Diarra:

L. velutina is used against a kind of dermatosis: small boils all over the body that are

itching (could be scabies).The bark is used for preparation of a decoction. It is used as

bath and is also drunk. If the skin infection start with dysenteria and the decoction is

drunk in the beginning, the dysenteria will stop. Drink half a cup (200ml). For boys less

than 4 years use approximately 50 ml. Used for all ages.

Bourama Traoré:

The plant is against a skin disease that starts with small boils. A liquid then forms, and

when the boils open the liquid will make new boils. This is dangerous for children. Take

trunk bark and divide it in two. The first half is pound and dried to a fine powder, the

second half is used to make a decoction. Use the latter for a bath and apply the powder on

the boils afterwards.

Konimba Traoré:

The stem bark is powdered and applied on the wound. The wound is first washed with

water and is healed within a week.

N`Pankourou Fané:

L. velutina is used against a disease that is called Sainiama n`kononiama ntorianama.

This gives convulsions with fever. Take the leaves and the stem bark and make a

decoction. Take a bath and drink the decoction, this is done 2-3- times per day.

23

Djenfa Koumaré:

Lannea velutina is used against chronic wounds. Dry the trunk bark, make a powder,

wash the wound and apply the powder on the wound once per day.

Uses against dermatosis:

Bourama Toure is using the bark from the plant on black dermatitis; it appears on the skin

in a round shape and itches in the beginning. The bark is dried, pound to a fine dry

powder, mixed with oil and applied on the skin.

The same healer has also another recipe for the same black dermatosis.

When the dermatosis itches a lot, the bark of Pterocarpus erinaceus and Afzelia africana

is used to make a powder. This powder is mixed with the powder of the bark of L.

velutina. One part is used for making a decoction that is used to wash the skin and the

other part is mixed with oil and applied on the dermatosis once per day until it is

completely cured.

Didieni-region, 6.2.04

Doneke Traore:

The trunk bark is used for the preparation of a decoction. The decoction is used for a bath

once per day to treat small pox.

Diotin Traore:

A decoction of trunk bark is used against wounds. Powder is applied on the wound after

washing with the decoction.

Balla Sissoko:

L. velutina is used against female infertility. Powder of the bark is mixed with water and

drunk

Konimba Keita:

A decoction of the stem bark is used as a bath to treat small pox.

Tietin Coulibaly:

The decoction of the trunk bark is used as a bath against itching

Cho Fane:

A decoction of the bark is drunk and also used as a bath against itching caused by small

boils on the body.

Massantola-region, 7.2.04

Fah Diarra:

L. velutina is used against dysenteria, but only if there is blood in the faeces. Make a

decoction of the rootbark and drink this.

24

Soundje Coulibaly:

The plant is used against gastric ulcer. A powder of the trunk bark is mixed with salt and

eaten.

N`tiokon Traore:

A decocotion of the trunk bark is made and used to treat small boils on the body. It is

used for washing the body or as a bath. The local name for the disease is “Niama”.

Soungalo Traore:

Powder of the stem bark is applied on wounds.

N’gogna Coulibaly:

A decoction of powdered trunk bark is used against dysenteria. The decoction is drunk

morning and evening.

Dossere Diarra:

L. velutina is used against inflammation. The juice of the trunk bark is applied on the

inflammation.

Naoba Diabati:

A decoction of the trunk bark is used as a bath against itching (small boils on the body).

Baba Diarra:

The plant is used against head ache. A decoction of the leaves is used to take a vapour

bath and to wash the head.

Deese Coulibaly:

A decoction of the leaves is used for washing the eyes to treat conjunctivitis.

H’toriké Coulibaly:

L. velutina is used on small boils. A decoction of the leaves is used to wash the area.

Powder of trunk bark is then applied.

N’kodjiri Traoré

Dysenteria and haemorrhoids; a decoction of the stem bark is drunk.

Stem bark and/or leaves were the plant parts used by all the interviews healers. 14 of the

healers use L. velutina to treat different skin diseases, from wounds to small pox. 3 use it

against dysenteria, especially with blood in faeces. It is also used to treat head ache,

conjunctivitis and haemorrhoids. 3 of the interviewed healers did not use this plant.

A search using www.google.com gave 40 hits and www.PubMed.com gave only 1 hit

(25.02.2005).

Sory Diallo is currently examining several Lannea species. The results can be found in a

thesis that should be finished by the end of 2004 or the beginning of 2005.

25

3.1.2.3 Mangifera indica L.

Thesis: Aouissa (2002)

Some traditional uses in Senegal:

Different astringent preparations made from the bark and leaves are used against

diarrhoea and dysentery. It is often recommended to eat the young leaves with a “white

kola nut”. The leaves are said to be used in the treatment of mental disorders. (Kerharo

and Adam, 1974). The fruits, mangos, are also widely known and used especially as food.

Aouissa (2002):

This author studied toxicity and other pharmacological properties of M.indica with the

purpose of using this plant in an ITM. Leaves from Mangifera indica have been used

combined with Fagara zanthozylodes and Securidaca longependunculata to treat mouth-

and teeth disorders (Aouissa).

The plant material was collected in the “Quartier du fleuve”-district in Bamako in

November 2001. It was dried in shadow and crushed to a fine powder when dry.

All animal tests were performed on female and male mice from Oncins France Souche 1

(OF1) with a mass between 19 and 25 g.

Extractions:

Drug 250 g

/ Petrol ether

___________________ / \

Extract with petrol ether \

Residue

/ DCM

_______________________/ \

Extract with DCM \

Residue

/ Methanol

________________________/ \

Extract with methanol \

Residue

/ H2O 50 º C

___________________________/ \


Residue

/ H2O 100 º C

_____________________________/ \



26

In addition to this, a maceration and a decoction with water were made.

A liquid-liquid separation was performed with 30 g of the lyophilized solution from the

decoction. The liquids used were petrol ether, ethyl acetate and butanol.

Tests:

Standard phythochemical tests (A.1.1) were performed earlier at DMT.

Thin layer chromatography was performed with all the extracts by using one of the

following solvent systems:

Water and methanolic extracts: Ethyl acetate:methyl ethyl ketone:formic acid:water

(50:30:10:10).

Petrol ether and DCM extracts: Ligroine:ethylacetate (2:1).

All plates were treated with Godins reagent and examined using UV light at 254 and 366

nm.

The total content of polyphenolic compounds in the drug was measured.

The antioxidant properties of the extracts were tested by using thin layer chromatography

combined with DPPH.

Antifungal activity was tested using the agar-overlay method

Haemostatic activity was tested by measuring coagulation time on full blood and

recalcification of blood plasma.

Toxicity: LD50 was determined after oral and intraperitoneal administration.

Analgetic effect was tested using “Hot plate” and “Writhing” tests.

Anti-inflammatory effect was tested using Carrageenan oedema test.

Results:


Flavonoids, tannins, mono- and polysaccharides, mucilages, sterols and/or triterpenes,

leucanthocyanines and cardiac glycosides. The content of water was between 5,18 and

5,60 %. Ashes insoluble in HCl were 2,52 %, total ash content was 4,88 % and sulphur

containing ash was 5,83 %.


A spot with Rf = 0,55: intense yellow/orange fluorescense at 366 nm, intense yellow

color with Godin at 254 nm. Spots with Rf = 0,55; 0,6; 0,66; 0,72; 0,8 and 0,85 are

visible at 254 and 366 nm in the polar solvents. They have a brown to purple

fluorescence. Some are colored with Godin. This indicates the presence of flavonoids in

the extracts.

27

Spots with Rf = 0,13 and 0,2 with blue to brown fluorescence at 366 nm is found in all

extracts except the ethyl acetate extract. The blue color might indicate the presence of

coumarins in the polar extracts. The results from the thin layer chromatography

correspond well with the result from the phytochemical tests. Most of the spots were

found in the polar extracts. This means that most of the substances are water soluble.

Content of polyphenolic compounds:

The content of polyphenols was calculated by using a standard curve. It was found to be

106,8 mg per 1 gram dried plant powder.

Antioxidant activity:

Table 3.1.2.3.a Results from tests on antioxidant activity

Extract Number of spots

Water (decoction?) 5

Ethyl acetate 8

Buthanol 4

Saturated water, water at 50 ºC and at

100ºC

1

Water maceration 3

Methanol 7

Fungistatic activity: no activity was observed.

Table 3.1.2.3.b Haemostatic activity

Extracts Coagulation time for blood (minutes)

Tube Drop Tube Drop Tube Drop Tube Drop

10 µl 25 µl 50 lµ 100 µl

Negativ control 12,38 12,07 12,38 12,07 12,38 12.07 12,38 12.07

Decoction 17 --- 4,3

(65 %)

5,3

(56 %)

--- 10,30

(14 %)

H H

Petrol ether 7

(43 %)

--- 8

(35 %)

9

(25 %)

7

(43 %)

14 10

(19 %)

---

Saturated extract 6,30

(49 %)

H = haemolytic activity was observed instead of haemostatic activity.

28

Table 3.1.2.3.c Plasma recalcification time

Test number Negative control Decoction (10 µl)

1 3,30 2,30

2 3,30 2,30

3 3,30 2,30

4 3,30 3,00

5 4,30 3,30

6 5,00 3,30

7 5,30 4,00

8 6,00 4,00

9 5,30 4,00

10 5,30 4,00

Mean 4,44±1,06 3,25±0,75*

Percentage reduction: 26,8 %

Student t test: *P < 0,0001 (very significant).

Coagulation time for milk:

The test was positive. This, and the positive results from the other tests on haemostatic

activity, might be caused by tannins in the leaves. Tannins have astringent properties that

might cause vasoconstriction and thereby influence the haemostasis.

LD50 after oral administration:

No deaths were observed. LD50 is above 3 g extract/kg body weight from lyophiliate and

hence a bigger LD50 than crude drug. The lyophiliate from the decoction of the leaves

from Mangifera indica is not toxic after administration of single dose taken orally.

Table 3.1.2.3.d LD50 after intraperitoneal administration:

Dose mg/kg Tested mice Dead mice % dead mice

500 6 0 0

750 6 0 0

1000 6 1 16,67

1250 6 5 83,33

1500 6 6 100

LD50 was calculated from the TREVAN curve to be 1100 ± 102, 10 mg/kg body weight.

Considering the extraction yield this gives a LD50 of 6,16 g/kg body weight for the crude

drug.

29

Analgetic activity:

Table 3.1.2.3.e Results from “Hot plate” test

Treatment Dose

(mg-

mg/kg)

Before

administration.

After administration. Reaction time in seconds

and % increase in reaction time.

10 min 30 min 60 min 90 min 120 min

Water 25 7,35 ± 1,63 3,24±1,20 5,17±0,06 7,55±3,20 3,78±1,89

M.indica 1000 2,75 ± 0,82 3,50±1,54 2,22±0,64 2,15±1,18 2,57±1,03

Tramadol 5 4,17 ± 0,81 18,3±4,2

338,84 %

24,67±4,70

491,6 %

22,18±6,49

431,89 %

9,88±3,79

136,93 %

The extract shows no effect on CNS except a slight reduction in reaction time compared

with the negative control test. The author believes this might be caused by saponins in the

plant.

Table 3.1.2.3.f Results from “Writhing” test

Treatment Dosage ml or

mg/kg

Number of

”torsions” mean

and sd

% inhibition

Distilled water 25 71,17 ± 6,49 ---

M. indica 500 48,17 ± 8,37* 32,32

M. indica 1000 46,53 ± 9,85* 34,19

M. indica 1500 41,33 ± 7,37* 41,92

Indometacin 25 mg 5 33,67 ± 3,5* 52,69

Tramadol 5 30,50 ± 6,71* 57,14

*P<0,01 Student t test

The extracts give some protection against chemically induced pain. The effect is

comparable to indometacin (5 mg/kg). The analgesic effect is dose dependent.

Table 3.1.2.3.g Anti inflammatory activity


mg/kg

Mean ± SD % inhibition

Distilled water 25 0,117 ± 0,008 ---

M. indica 250 0,094 ± 0,017** 19,86

M. indica 500 0,094 ± 0,017** 19,86

M. indica 1000 0,075 ± 0,028** 36,14

ASA 100 0,074 ± 0,010** 36,86

Indometacin 5 0,089 ± 0,013** 23,43

The mean is calculated for 6 mice. ** P<0,01 Student t test

All the extracts show anti-inflammatory effects in this test.

30

Table 3.1.2.3.h Prolonged anti inflammatory activity:


mg/kg

1 hour after

injection

3 hours after

injection

5 hours after

injection

Distilled water 25 0,093 ± 0,134 0,117 ± 0,008 0,091 ± 0,007

M. indica 1000 0,085 ± 0,021

(8,27 )

0,075 ± 0,028

(30,52 %)

0,092 ± 0,017

(- 0,36 %)

The anti-inflammatory activity is strongest after 3 hours. The activity after 1 hour is very

small, and no activity is seen after 5 hours.

All the tested extracts showed some antioxidant effect. This might be cause by the high

content of polyphenolic compounds (tannins and/or flavonoids) in the leaves from M.

indica. The large number of active spots in the water extracts supports the traditional use

of water decoctions from this plant.

All the tested extracts show some haemostatic activity but the activity is not dose

dependent. The decoction shows the strongest activity.

A search using www.google.com gave 42900 hits and www.PubMed.com gave only 110

hits (25.02.2005). For more information about this plant see Nguyen (2001).

3.1.2.4 Spondias mombin L.

Thesis: Sangare (2003)


In Senegal the juice from fresh leaves is used against eye diseases. A decoction made

from the leaves is used to treat dysenteric diarrhoeas, and a maceration from the roots is

used against painful colic. A decoction of leaves and bark is used to treat post partum

bleeding and to treat women who have had several abortions. (Kerharo and Adam, 1974).

Sangare (2003) This author tested different parts of 8 plants for activity against Plasmodium falciparum.

The plants were identified after interviews with several healers in Kendié and Finkolo in

January and February 2003. 49 healers were interviewed in Kendié and 30 in Finkolo.

They were asked which plants they used to treat malaria, and also what they believed

were the cause of the disease. In Kendié 22,45 % believed that mosquito bites caused

malaria, while the number was only 10 % in Finkolo.

Leaves from S. mombin were collected in Blendi in the Sikasso region in May 2003.

Extractions:

50 g plant material + water (500 ml – 1 hour)

/

Decoction

Figure 3.1.2.4.a Decoction

31


/

Water maceration

Figure 3.1.2.4.b Maceration with water

Plant material (50g)

/ DCM (2 x 24 hours)

_____________ / \

Extract with DCM \

Residue

/ methanol (2 x 24 hours)

________________________/ \


Residue

/ H2O 50 º C (1 hour)

___________________________/ \

Extract with H2O at 50 º C (digestion) \

Residue

/ H2O 100 º C (1 hour)

_____________________________/ \

Extract with H2O at 100 º C \ Residue

Figure 3.1.2.4.c Extractions with solvents of different polarity

Tests:

Standard phytochemical tests (A.1.1) were performed at DMT.

Activity against Plasmodium falciparum was tested.

Thin layer chromatography was performed on some of the decoctions and digestions.

Solvent system BAW (60:15:25). The plates were treated with AlCl3 and Godin’s

reagent.

Results:


Flavanoids, tannins, saponins, anthraquinones, coumarins, reducing compounds, mono-

and polysaccharides and mucilages. The content of water was between 7 and 9,56 %.

Ashes insoluble in HCl were 0,94 %, total ash content was 4,24 % and sulphur containing

ash was 4,02 % (Table 3.2).

The thin layer chromatography showed two spots that probably correspond with content

of flavonoids.

32

IC50-concentrations:

DCM extract: 2,57 µg/ml

Methanol extract: 3,92 µg/ml

Maceration with water: 7,66 µg/ml

Decoction: 7,89 µg/ml

Chloroquine: 0,05 µg/ml (reference substance)

All extracts from the leaves of S. mombin showed good activity against P. falciparum.

For more information about this plant see Diallo et al (2002) and Tran (2004).

3.1.3 Annonaceae

3.1.3.1 Annona senegalensis Pers.

Theses: Kanta (1999) and Samake (2000)


This medicinal plant is believed to have magical and medicinal properties, and it is

widely used because of this. The most common uses are against diarrhoea and dysentery,

and the most common formulation is a maceration of the bark from the branches or small

branches with leaves or a powder dispersed in liquid (for example milk). The leaves and

roots are said to be: antipyretic, antitussive, antiseptic, diuretic, anti-infective and to have

several other medical properties. They have been used to treat the following diseases:

malaria, respiratory diseases, eye diseases, dermatosis, rheumatism and other diseases.

This plant might be used alone or combined with other plants (Kerharo and Adam, 1974).

Kanta (1999) This author identified 29 plants used in the treatment of infections by Candida albicans,

These were identified after interviews of healers and salesmen of medicinal plants and a



of

November 1998 in the Tienfala forest, 30 km from Bamako and 10th

of December 1998

in mount Manding in Siby, 60 km from Bamako, dried, pulverized and later analyzed.

Extractions:

Leaves, stem bark and root bark from A. senegalensis was analyzed. Extracts were made

with ethanol, dichloromethane and methanol.

Tests:

All extracts were tested for activity against Candida albicans and Escherichia coli.

33

Results:

None of the extracts had effect against E. coli. The ethanol extract from the root bark and

the dichloromethane extract from the stem bark showed effect against C. albicans at 30

mg/ml. Activities: ++ and +++ (maximum activity is marked with +++).

At 10 mg/ml, the dichloromethane extract of the stem bark gave 7 active products using

thin layer chromatography plates combined with the same microorganism. The ethanol

extract from the root bark gave 3 products using the agar overlay method.

Samake (2000)




Siby in November 1998, and all plant material was dried and pulverized before analyzis.

Extractions:


/ Ethanol (80º)

________________________/ \


Residue

/ H2O 50 º C

___________________________/ \


Residue

/ H2O 100 º C

_____________________________/ \


Figure 3.1.3.1.a Extraction with solvent of different polarity

A dialysis was performed on all solutions, followed by lyophilization.

Tests:


All solutions were hydrolyzed before testing to separate polysaccharides into separate




Gas chromatography was also used to determine the monosaccharide content in the

different plants. Standard here was mannitol.

Activity on the complement system was tested on sheep erythrocytes. ICH50 is the

concentration that gives 50 % inhibition lysis of sensible erythrocytes.

34

Results:


Table 3.1.3.1.a Extracts tested with known sugars as references and ICH50

Tested

substance

Tested

part


(µg/ml)

A.s. Leaves + + + 0 + + + + 0 > 500

A.s. “ + + + 0 + + + + 0 150

A.s. Root

bark

+ + + 0 + + + + 0 25

A.s. “ + + + 0 + + + + 0 130

A.s. Stem

bark

+ + + 0 0 + + + 0 125

A.s. “ + + + 0 0 + + + 0 25

A.s. = Annona senegalensis

Table 3.1.3.1.b Amounts of monosaccharides and polysaccharides detected

Leaves Root bark Stem bark

Monosaccharides Extract at

50ºC

Extract at

100ºC

Extract at

50ºC

Extract at

100ºC

Extract at

50ºC

Extract at

100ºC

Sugar

ratio (%)

Sugar

ratio (%)

Sugar

ratio (%)

Sugar

ratio (%)

Sugar

ratio (%)

Sugar

ratio (%)

Arabinose 8 3 28 3 1 5

Rhamnose 29 35 9 1 1 2

Xylose 2 0,5 48 3 2 36

Mannose 1 0,5 4 1 0 0

Galactose 16 17 11 1 2 5

Glucose 7 3 11 92 2 27

Glucuronic acid 0 0 0 0 14 20

Galacturnoic

acid

37 42 13 0 51 5

Total 100 101* 124* 101* 73* 100

Polysaccharides

content

53 % 32 % 44 % 58 % 60 % 35 %

*The only information availabe is a summary table – basic data needs to be checked.

ICH50 (µg/ml) values for 5 of the 6 tested extracts is under 150. This might be cause by

the relatively high content of polysaccharides in all plant parts (32% to 60 %).

For more information about this plant see Aasberg (2001) and Diallo et al (2002).

35

3.1.4 Araliaceae

3.1.4.1 Cussonia arborea Hochst. ex. A. Rich. (Synonym: Cussonia barteri

Seem.)

Theses: Bouare (2004)and Ouologuem (1999)


Strømland (Strømland 2003) interviewed 32 healers in the Dogonland in Mali. The main

uses identified from the interviews were: stomach pain, oedema, Hansen’s disease

(leprosy), mental diseases, constipation, sexually transmittable diseases and flatulens.

Bouare (2004) This author identified traditional use of Cussonia arborea, and recipes based on this

plant, by interviewing traditional herbalists/healers in the Dioila area in Mali in February

2003. According to these interviews the following uses were found:

Visual troubles, gonococci, oedema, syphilis, indigestion, stomach ache, toxoplasmosis,

Hansen’s disease (leprosy), food intoxication, head ache, asthma, to help during child

birth and painful menstrual periods, wounds, muscular pain, blackouts, swelling (non

specific), bloated stomach and dermatosis.

The plant material was collected in Blendio in the south of Mali. Only the bark from the

root was analyzed.

Extractions:


/

Decoction



/

Water maceration


50 g plant material + ethanol (500 ml x 24 hours x 3)

/

Ethanol extract

Figure 3.1.4.1.c Maceration with ethanol

36

200 g plant material + dichloromethane (DCM) (1500 ml x 24 hours x 3)

/

DCM extract

Figure 3.1.4.1.d Maceration with DCM

Tests:



combined with DPPH.

Thin layer chromatography was performed, using the following solvent systems: -

Buthanol:acetic acid:water – 60:15:25 (BAW) for water and ethanol extracts.

Ligroine:ethyl acetate – 1:1 for the DCM extract. The thin layer plates were treated with

Godins reagent and examined using UV light at 254 and 366 nm.

Larvicidal activity:

Test on Anopheles gambiae and Culex quinquefasciatus.

Cytotoxic activity:

Test on Lombricus terrestris was performed to measure cellular toxicity.

Molluscicide activity was tested on Bulinus truncatus and Biomphalaria pfeifferi.

Antibacterial activity was tested on the following organisms:

Acinetobacter calco var anitrat

Citrobacter freundii

Enterobacter agglomerans

Escherichia coli

Klebsiella penumoniae

Proteus mirabilis

Pseudomonas aeruginosa

Salmonella spp

Streptococcus β haemolytic

Antifungal activity was measured using the Agar overlay method.

Results:


Flavanoids, leucoanthocyans, tannins, saponins, cardiac glycosides, sterols and/or

triterpenes, anthraquinones, coumarins, mono- and polysaccharides and mucilages. The

content of water was between 5,37 and 8 %. Ashes insoluble in HCl were 2,8 %, total ash


The result from the thin layer chromatography corresponds with the results from the

phytochemical tests.

37

Only a modest antioxidant activity was observed in the water and ethanol extracts.

This corresponds well with absence of reducing compounds in the phytochemical tests.

All extract had larvicidal effect on Anopheles gambiae. The DCM extract showed the

strongest activity with 100 % deaths at a concentration of 250 ppm (0,025 mg/ml).

Only the DCM extract showed effect on Culex quinquefasciatus. A concentration of 500

ppm (0,05 mg/ml) gave 95 % deaths.

Only the ethanol and DCM extract showed effect on Lombricus terrestris. The ethanol

concentration gave 100 % deaths at concentration of 3,75 mg/ml while the DCM extract

gave 100 % deaths at 1,875 mg/ml.

All extracts showed effect on Bulinus truncatus and Biomphalaria pfeifferi at a

concentration of 400 ppm (0,04 mg/ml).

Bulinus truncatus: the water and DCM extracts gave 100 % deaths at this concentration

while the ethanol extract gave 100 % deaths at 200 ppm (0,02 mg/ml).

Biomphalaria pfeifferi: the decoction gave 100 % deaths at 400 ppm (mg/ml), the DCM

extract gave 100 % deaths at 200 ppm (mg/ml) and the ethanol extract and the water

maceration gave 100 % deaths at 100 ppm (mg/ml).

Table 3.1.4.1.a Antibacterial tests

Measurement of inhibition zones in mm

Species DCM DCM EtOH EtOH M

H2O

M

H2O

D H2O D H2O

3 µl 4 µl 3 µl 4 µl 3 µl 4 µl 3 µl 4 µl

S.aureus 6,5 7 6,5 7 8 8 6,5 8

Streptococcus 7 7 7 7 7 7 7 9

C.freundii 8 8,5 7 7 0 0 7 9

E.coli 8 8 0 0 0 0 0 0

K.pneumoniae 7 7 6,5 6,5 7 7,5 7 7,5

E.agglomerans 8 9 9 9 6,5 7 7 8

A.c var antirat 6,5 6,5 9 9,5 7 7 8 9

S.spp 0 0 0 0 0 0 7 9

P.mirabilis 7 7,5 0 0 0 0 7 7

P.aeruginosa 0 0 0 0 0 0 0 0

The decoction showed best antibacterial activity.

Table 3.1.4.1.b Antibacterial activity of reference species

Measurement of inhibition zones in mm

Reference

species

DCM DCM EtOH EtOH M

H2O

M

H2O

D H2O D H2O

3 µl 4 µl 3 µl 4 µl 3 µl 4 µl 3 µl 4 µl

S.aureus 10 10 6,5 7 8 8 7 8

E.coli 8 8 0 0 6,5 7 7 7

P.aeryginosa 0 0 0 0 0 0 0 0

38

Antifungal activity: the DCM extracts gave inhibition zones for 100 µg, 200 µg and 300

µg extracts at Rf 0,93. The diameter of the zones varied with the tested concentration.

Ouologuem (1999)

This authour tested the larvicidal effects of different extracts from 11 Malian plants.

Larvae from Anopheles gambiae and Culex quinquefasciatus were used. 6 of the plants

were tested on both species, while 5 were only tested on C. quinquefasciatus.



Darsalam, in the Bamako district.

Pre-made extracts were used. DCM, methanol and water extracts from C. arborea roots

were tested.

Tests:




tests were performed on 5x20 larvae, and larvae that had fallen to the bottom of the tubes

or were immobile were considered dead.





Anopheles gambiae.

Results:

Larvicidal tests:

Table 3.1.4.1.c Culex quinquefasciatus - % deaths at 500 ppm

Plant extract

Time of exposure C.a. H2O C.a. Methanol C.a. DCM

30 minutes 10 10 25

1 hour 10 15 65

24 hours 60 20 100

Table 3.1.4.1.d Anopheles gambiae - % deaths at 500 ppm

Plant extract

Time of exposure C.a. H2O C.a. Methanol C.a. DCM

30 minutes 90 75 55

1 hour 90 80 90

24 hours 95 100 100

C.a. = Cussonia arborea

39

Extended tests at different concentrations: See tables 3.1.2.2.c - 3.1.2.2.f.

This plant shows good antibacterial, antifungal and larvicidal effects.

Strømland (2003) isolated and characterized lowmolecular substances from this plant.

See his thesis for more information about C. arborea.

Cussonia arborea:

A search using www.google.com gave 95 hits and www.PubMed.com gave 1 hit

(25.02.2005).

Cussonia barteri:

A search using www.google.com gave 76 hits and www.PubMed.com gave 3 hits

(25.02.2005).

3.1.5 Arecaceae

3.1.5.1 Borassus flabellifer L.

Traditional uses:

In Senegeal, preparations made from roots of young plants are used to treat sore throat

and bronchitis.Palm wine is made from sap and is consumed by non-Muslims as a

stimulating agent. It is also a part of most of the aphrodisiac remedies made (Kerharo and

Adam, 1974).

Field work:


N`Tjobougou, 5.2.04

Djokin Diarra is using the flower from this plant against leaking boils. The flowers was

mixed with Fomes fomentarius (English: german tinder. Norwegian: kjuke) and

carbonized. The powder was mixed with butter from Butyrospermum parkii, arachis or

palmtrees.

Bourama Toure uses the flower from this plant against black dermatosis; this appears on

the skin in a round form and itches in the beginning. The flowers are carbonized into a

powder and mixed with butter from Butyrospermum parkii before it is applied.

Konimba Traore is using the flower from B. flabellifer against dermatosis on the head.

Carbonize the flower, mix the powder with oil and apply the mixture on the dermatosis.

Didieni-region, 6.2.04 and Masantola-region, 7.2.04

Donéké Traoré is using the flower from this plant to treat black dermatosis. Dry the

flower, carbonize it and mix the powder with the butter from B. parkii. Apply 2 times per

day.

40

Diotin Traoré uses the roots from this plant to treat black dermatosis. The flower is

carbonized, mixed with the butter from B. parkii. The mixture is applied once.

Soungalo Traoré, N`gogna Coulibaly, Naoba Diabati and N`kotjiri Traoré are using the

flower from this plant to treat dermatosis. Carbonize the flower, mix with the oil of B.

parkii and apply on the body.

H`toriké Coulibaly uses the flower from B. flabellifer to treat dermatosis. Carbonize the

flowers, mix with potash and oil from B. parkii and apply on the skin.

10 of the 21 interviewed healers use the carbonized flower from this plant to treat

dermatosis.The plant is a palm like tree, and the flowers are quite big.

Further examination of their phytochemical content and other properties might be

interesting.


(25.02.2005).

3.1.6 Asclepiadaceae

3.1.6.1 Calotropis procera (Aiton) W.T. Aiton


In Senegal the plant is known to be toxic. The latex, leaves and especially the roots are

emetic. It is therefore used as a purgative and emetic remedy and also against poisoning.

The root bark is used in the treatment of anxiety and other mental disorders.

The roots are used as a part of a treatment against syphilis and Hansen’s disease

(leprosy). In all cases the preparations contains only small amounts of the plant since the

cardio toxic effect is normally well known. The latex is used externally as antiseptic

agent and as a sedative (Kerharo and Adam, 1974).

Field work



Donéké Traoré is using the stem from this plant to treat black dermatosis. Dry the stem,

carbonize it and mix the powder with the butter of B. parkii. Apply 2 times per day.

Balla Sissoko uses the stem and leaves from C. procera to treat itchy, black dermatosis.

Make a decoction and use as a bath.

Konimba Keita uses the leaves from this plant to treat dermatosis. The leaves are

carbonized, mixed with the butter from B. parkii. This is applied to the skin once daily.


(25.02.2005).

41

3.1.6.2 Leptadenia hastata (Pers.) Decne.


The easy access to this plant is probably the main reason for its common use.

The latex is used to treat wounds, while a maceration of the roots are used against

constipation. The whole plant is used to treat vaginal discharge, while the leaves are used

to treat weakness and problems with lactation. Different parts of the plants are used in

combination with other plants to treat syphilis, urinary problems, Hansen’s disease,

constipation, mental diseases and diarrhoea (Kerharo and Adam, 1974).

Field work:

Dideni-region, 6.2.04

Diotin Traoré uses the flower from this plant to treat black dermatosis. The flower is

carbonized and mixed with the butter from B. parkii. The mixture is applied once.


(25.02.2005).

3.1.6.3 Leptadenia pyrotechnica (Forssk.) Decne.

Thesis: Bah (1998)


In Senegal, Leptadenia sp. has been used as a laxative for children that are less than 10

days old. It is also used against pain in the kidneys and sore muscles (Kerharo and Adam,

1974). In Mali, the stem juice is administered twice a day for 4 or more days in the

nostrils to cure cough, colds and running nose (Diallo et al, 1999).

Bah (1998)




new and possibly less toxic agents from plants to try and control the disease. Anopheles

gambiae mosquitoes were collected in Mopti. For the tests, larvae were elevated and

collected in the laboratory, from full grown females collected at one specific date. Larvae

were fed with Whiskas ® cat food.

The plant material was collected in Hombori in June 1996, and was dried and pulverized

before analysis. Only the branches from L. pyrotechnica were analyzed.

An extract was made with water, 500 ml x 24 hours x 3. The extract was tested at

concentrations from 0,015 mg/ml to 0,06 mg/ml on groups of 8 x 25 larvae and the

number of dead larvae registered. No deaths were observed at any of the tested

concentrations.

42

The branches from this plant do not show any activity in the performed tests. This plant

will not be discussed further in this thesis.

A search using www.google.com gave 358 hits while www.PubMed.com gave no hits

(25.02.2005).

3.1.7 Asteraceae

3.1.7.1 Vernonia colorata (Willd.) Drake

Thesis: Fane (2002)


This plant is used by healers everywhere it is found. The leaves have a bitter taste that is

said to resemble quinine and they are used against fever and stomach aches. Inhalation of

decoction from leaves are used in epileptic seizures. It is also comined with other plants

in the treatment of schistostomatiosis and sterility (Kerharo and Adam,1974).

Fane (2002) This author interviewed 110 healers in the Bamako district; 50 men and 60 women. Most

were in the age group between 40-70 years. They were asked to name known toxic

plants, the use of these plants, toxic effects and possible anti-dotes.

299 examples were mentioned, giving a total of 61 different plants. Maiga et al (2004)

did a literature research of the 19 most cited plants.

Phytochemical tests were performed on 5 of these plants and, further biological tests were

performed on 3 plants: Vernonia colorata, Afrormosia laxiflora (3.1.19.1) and Cassia

siberiana (3.1.11.4). The plants were chosen from both the results from the interviews,

but also from the literature about the plants.

Table 3.1.7.1.a The 10 most cited plants (with indication for use) in the interviews:

1. Swartzia madagascariensis (39) Jaundice, malaria

2. Cassia sieberiana (23) Stomach ache, malaria

3. Securdiaca longepedunculata (20) Haemorrhoids, small pox

4. Trichilia emetica (19) Haemorrhoids, gastric ulcer

5. Cassia alata (17) Constipation, malaria

6. Entada africana (12) Malaria

7. Anogeissus leiocarpus (12) Constipation, malaria

8. Gardenia ternifolia (10) Jaundice, malaria

9. Acacia senegal (10) Vulvovaginitis, haemorrhoids

10. Opilia celtidifolia (8) Malaria, stomach ache

43

Table 3.1.7.1.b The 10 most cited indications were:

1. Malaria

2. Stomach ache

3. Constipation

4. Jaundice

5. Haemorrhoids

6. Vulvovaginitis

7. Gastric ulcer

8. Vaginal discharge

9. Migraine

10. ”Mauvais sort”

Vernonia colorata leaves were collected from the Mambilé school-garden in November

2001. The plant material was dried and pulverized before use.

Other biological material:

- Mice of the strain “Oncins France Souche 1” were used. Their weight was

between 20-25 g.

- Refrigerated blood from cattle was collected at the slaughter house in Bamako

in March 2002.

- Anopheles gambiae larvae (stadium 2) were acquired from the “Department

de l’epidemiologi des affections parasitaires” in Bamako.

- Full grown Lombricus terrestris were collected by the river bank in Djoliba in

June 2002.

Extractions:

A decoction was made with 250 g powdered plant material in 2,5 l distilled water for 3

hours.

A maceration was made with 50 g powdered plant material in 500 ml distilled water for

3x24 hours.

Tests:


Toxicity:

LD50 orally was tested on groups of 6 mice (3 male and 3 female). Dosage of extracts was

from 500 to 5000 mg/kg. Mortality was observed until 3 days after administration.

LD50 intraperitoneal was tested on groups of 6 mice (3 male and 3 female). Dosage of

extracts was from 400 to 675 mg/kg. Mortality was observed until 3 days after

administration.

Chronic/prolonged toxicity was tested on groups of 16 mice (6 male and 10 female).

Extracts were administration orally at a concentration of 42,75 mg/kg body weight for 4

weeks. After 2 weeks, 2 mice from each group were killed and their organs were

examined. After the 4th

week, the rest of the mice were killed and their organs examined.

44

Transaminase test.

Haemolytical tests.

Larvicidal activity.

Test of cytotoxic effect on Lombricus terrestris.

Results:


Alkaloids (0,11 %), flavonoids, tannins, saponins, coumarins, reducing compounds,

mono- and polysaccharides, mucilages, sterols and/or triterpenes, leucanthocyanines and

cardiac glycosides. The content of water was between 5,65 and 6 %. Ashes insoluble in

HCl were 10,27 %, total ash content was 13,6 % and sulphur containing ash was 19,17

%.

Toxicity:

LD50 orally: no deaths observed.

LD50 intraperitoneal: no deaths observed, but the mice showed shaking and had

diarrhoea.

Chronic/prolonged toxicity: no deaths observed, but an increase in weight for both male

and female mice.

The weights of liver and kidneys from treated mice were comparable to the values for

mice that were only given water. One of the female mice got an enlarged pancreas.

Transaminase:

1. Serum from healty mice

2. Serum from mice that were given distilled water.

3. Serum from mice that were given Vernonia colorata

Table 3.1.7.1.c Results from transaminase test

Serum SGOT SGPT

1 34 26

2 65 38

3 97 34

Both SGOT and SGPT increased.

Table 3.1.7.1.d Results from larvicidal test on Anopheles gambiae

Extracts (500 µg) Number of larvae Time

24 hours

Number of deaths % dead

Distilled water 20 0 0

Decoction V.c. 20 16 80

Maceration V.c. 20 14 70

V.c = Vernonia colorata

No deaths were observed after 30 minutes or after 1 hour.

45

Table 3.1.7.1.e Results from Lombricus terrestris test:

Extract

90000 pp

Number

of L.t.

Mortality Number

of deaths

% dead

after 72

hours 24 hours 48 hours 72 hours

Distilled

water

5 0 0 0 0 0

Decoction

V.c.

5 0 0 0 0 0

Maceration

V.c.

5 5 - - 5 100

Haemostatic tests: negative

See Maiga et al (Maiga 2004) for more information about this plant.

3.1.7.2 Vernonia kotschyana Sch. Bip. ex Walp.

Theses: Ba (1998), Bah (1998) and Ouologuem (1999).

Traditional use:

The root of this plant is used in wound healing in Mali (Diallo et al, 2002). It is also used

in the treatment of gastric ulcer.

Ba (1998)

This author studied 6 plants traditionally used as “fishing poison” or used as insecticides

in cultivation of cereals, to determine the chemical composition and molluscicidal

activity of these plants. This was done to find plants that may be used in the prevention of

schistosomiasis by killing the molluscs that act as intermediate hosts.

Molluscs were collected in Djikoroni Woyowayemko and N’Gomidjirambougou. The

species used were Bulinus truncatus and Biomphalaria pfeifferi. Groups of 6 snails were

used in all tests. The plant material was found in the forest at Koulouba the 6th

of March

1997. Only the root bark was tested, and it was dried and pulverized before analysis.

Extractions: Drug 150 g

/ Acetone (500 ml)

___________________ / \

Acetone extract Residue

/ Ethanol (500 ml)

_______________________/ \

Ethanol extract Residue

/ Water (500 ml)

________________________/

Water extract

Figure 3.1.7.2

46

Tests:


Molluscicidal activity was tested for all extracts at concentrations from 0,025 mg/ml to

0,4 mg/ml. Tests were performed on Biomphalaria pfeifferi and Bulinus truncatus.

Results:



triterpenes, coumarins, reducing compounds, mono- and polysaccharides and mucilages.

The content of water was between 7,25 and 8 %. Ashes insoluble in HCl were 7,71 %,

total ash content was 8,74 % and sulphur containing ash was 12,21 % (Table 3.2).

Molluscicidal activity:

Only the acetone and ethanol extracts showed activity. A concentration of 0,2 mg/ml of

the acetone extract and 0,4 mg/ml of the ethanol extract 100 % deaths for Biomphalaria

pfeifferi,. A concentration of 0,2 mg/ml for the acetone extract gave 100 % deaths for

Bulinus truncatus, while the ethanol extract gave 50 % deaths at 0,4 mg/ml.

Bah (1998)








The plant material was collected in Sotuba in 1996, and was dried and pulverized before

analysis. Only the “tubercules” from V. kotschyana were analyzed.

Extractions were made with dichloromethane, methanol and water and all extracts were

tested on groups of 8 x 25 larvae at concentrations from 0,015 mg/ml to 0,06 mg/ml.

None of the extracts showed any activity.

Ouologuem (1999)

This author tested the larvicidal effects of different extracts from 11 Malian plants.

Larvae from Anopheles gambiae and Culex quinquefasciatus were used. 6 of the plants

were tested on both species, while 5 were only tested on C. quinquefasciatus.

Larvae from Culex quinquefasciatus were collected in N’Tomikorobougou and Darsalam,

in the Bamako district.

Pre-made extracts were used. Water, ethanol and acetone extracts from V. kotchyana

leaves were tested.

Tests:

Tests on larvae from Culex quinquefasciatus were done on 5,5 mg extract dissolved in

110 µl of a suitable solvent; water for the water and ethanol extracts and

47

dimethylsulfoxide (DMSO) for the methanol, ether and DCM-extracts. The tests were

performed on 5x20 larvae, and larvae that had fallen to the bottom of the tubes or were

immobile were considered dead.

Results:

Larvicidal test:

Table 3.1.7.2 Culex quinquefasciatus - % deaths at 500 ppm

Plant extract

Time of exposure V.k. H2O V.k. Ethanol V.k. Acetone

30 minutes 10 0 0

1 hour 10 10 10

24 hours 25 50 15

V.k. = Vernonia kotschyana

The tested extracts show molluscicidal activity and weak larvicidal activity.

Nergård et al (2004) have done a review of V. kotschyana and examined the

immunomodulating activities of polysaccharides found in the plant.


(25.02.2005).

3.1.8 Balanitaceae

3.1.8.1 Balanites aegyptiaca L.

Thesis: Ahamet (2003)

Some traditional uses in the Gourma district in Mali:

Root bark maceration is drunk to treat constipation and malaria. Fresh leaves crushed

with fruits of Acacia nilotica (L) Willd. Ex Del are mixed with cow’s butter and water

and the mixture is used to treat tonsillitis. Applied locally it may be used to treat gum-

inflammation. Fruit maceration is used against conjunctivitis. Almonds of crushed fruits

are dampened with water and laid on pimples. The almonds are also used to treat wounds

and scabies (Diallo et al 1999).

Ahamet (2003)

The authour studied this plant extensively.The plant material was collected in Niger in

the Maradi zone where the ground is sandy. The plant was dried in shadow for three

weeks before being ground to a powder in a traditional mortar. The stem bark of the plant

was analyzed.

Extractions were made with water (decoction, digestion and maceration), ethanol

(maceration) and chloroform (maceration).

48

Tests:

Thin layer chromatography was performed with all the extracts by using one of the

following solvent/migration systems:

Water and ethanol extracts: - Chloroform:acetone / 80:20

- Chloroform: acetone / 60:40

- Chloroform:methanol:water / 65:35:5

Chloroform extract: - Ligroine:ethylacetate / 2:1

The chromatography plates were treated with Godin and Dragendorff reagents and

studied using an UV lamp at 245 nm and 366 nm

The antioxidant properties of the extracts were tested using DPPH.


Biological tests: - Test for anti-inflammatory activity

- Test for analgesic activity

- Test for antispasmodic activity

Results:


Alkaloids (0,13 %), leucanthocyanines, tannins, saponins (abundant), cardiac glycosides,

sterols and/or triterpenes, coumarins, carotenoids, reducing compounds, mono- and

polysaccharides and mucilages. The content of water was between 4,86 and 6 %. Ashes

insoluble in HCl was 1,94 %, total ash content was 7,53 % and sulphur containing ash

was 11,18 % (Table 3.2).


Saponines and triterpenes gave a violet coloration with Godin’s reagent which

corresponds well with the results from the phytochemical tests.

In addition to this orange fluorescence at 366 nm and intense yellow coloration with

Godin’s reagent indicates the presence of flavonoids, despite their absence in the


Table 3.1.8.1.a Test for anti-inflammatory activity Treatment Dose

mg/ml/kg

M ± SD

M is mean volume of the mouse feet at different times

60 min 120 min 180 min 240 min

Negative control 10 1,49 ± 0,17 1,88 ± 0,42 2,15 ± 0,53 2,22 ± 0,44

Indometacine 10 1,13 ± 0,12** 1,15 ± 0,16** 1,18 ± 0,16** 1,32 ± 0,18**

Digestion H2O 300 1,17 ± 0,25* 1,28 ± 0,30* 1,37 ± 0,32* 1,48 ± 0,29**

Digestion H2O 900 1,03 ± 0,15** 1,20 ± 0,25* 1,29 ± 0,25* 1,29 ± 0,21**

Maceration H2O 300 1,25 ± 0,15* 1,23 ± 0,16** 1,25 ± 0,18* 1,33 ± 0,19**

Maceration H2O 900 1,15 ± 0,04* 1,19 ± 0,11 1,35 ± 0,13* 1,46 ± 0,19**

Ethanol 300 1,20 ± 0,07* 1,39 ± 0,15 1,55 ± 0,24* 1,70 ± 0,30

Ethanol 900 1,32 ± 0,25 1,33 ± 0,17 1,51 ± 0,22* 1,72 ± 0,28

Dichloromethane 300 2,29 ± 0,11** 1,34 ± 0,20 1,40 ± 0,18* 1,69 ± 0,20*

Dichloromethane 900 1,46 ± 0,67 1,33 ± 0,12 1,43 ± 0,12* 1,84 ± 0,08

49

M = mean volume of 5 rats. SD = standard deviation.

Student t test: *P < 0,05 = signifiacant **P = < 0,01 = very significant

All the extracts except dichloromethane at 300 mg/ml/kg at 60 minutes showed some

effect compared to the non-treated reference group. Digestion at a concentration of 900

mg/ml/kg showed the best effect after 60 minutes. The results show no dose dependence.

The animals used in this test were rats of race “Wistar” with a weight between 150 and

180 g.

Table 3.1.8.1.b Test for analgesic activity “Writhing test”

Treatment Dose mg/ml/kg Number of torsions

M ± SD

% Inhibition

Distilled water 25 48,00 ± 6,50 -

Paracetamol 100 32,00 ± 7,00** 33,33

Digestion H2O 100 40,20 ± 10,18 16,25

Digestion H2O 300 39,40 ± 5,54* 17,91

Digestion H2O 900 5,60 ± 5,07** 88,33

Maceration H2O 100 25,20 ± 7,59** 47,50

Maceration H2O 300 25,80 ± 6,76** 46,25

Maceration H2O 900 23,20 ± 4,02** 51,66

Decoction H2O 100 42,00 ± 6,00 12,50

Decoction H2O 300 25,40 ± 7,12** 47,91

Decoction H2O 900 21,40 ± 5,17** 55,41

Ethanol 100 32,40 ± 2,10** 31,66

Ethanol 300 30,80 ± 7,98** 35,83

Ethanol 900 31,60 ± 7,30* 34,16

M = mean volume of 5 mice. Student t test: *P < 0,05 = significant **P = < 0,01 = very

significant. SD = standard deviation.

The animals used in this test were female mice of the strain Oncins France Souche1.

Table 3.1.8.1.c Test for antispasmodic activity

Treatment Injected volume

(ml)

Concentration

(µg/ml)

% Inhibition

Atropine 0,15 0,0075 53,12 ± 4,7

Decoction H2O 0,2 100 22,22 ± 2,6**

Decoction H2O 0,2 300 44,60 ± 6,9**

Decoction H2O 0,2 900 54,84 ± 5,7**

Maceration H2O 0,2 100 16,66 ± 1,9 **

Maceration H2O 0,2 300 22,08 ± 2,4**

Maceration H2O 0,2 900 30,46 ± 3,4**

Dichloromethane 0,2 100 14,44 ± 1,9**

Dichloromethane 0,2 300 49,87 ± 4,2**

Dichloromethane 0,2 900 64,94 ± 3,68*

M = mean volume of 5 rats. SD = standard deviation.

Student t test: *P < 0,05 = significant **P = < 0,01 = very significant.

50

The animals used in this test were rats of race “Wistar” with a weight between 150 and

180 g. Most of the tested extracts showed analgesic activity. This activity might be cause

by saponins.

Numerous compounds showing antioxidant activity were found in all the extracts.

Substances like tannins, coumarins or carotenoids might be responsible for this activity.

The anti-inflammatory effect might be caused by saponins and/or triterpenes.

All the tested extracts showed good antispasmodic activity. This might be caused by

coumarins or alkaloids.

Reviews of this plant were done by Aasberg (2001).

3.1.19 Bignoniaceae

3.1.9.1 Stereospermum kunthianum Cham.


Traditional use:

In Mali, the powder of the dried bark is applied on burns or wounds. The wound is

washed with a decoction of fresh leaves. Special indications for sexual wounds (Diallo et

al, 2002).

Kanta (1999)


These were found after interviews of healers and salesmen of medicinal plants and a



of November 1998 in the Tienfala forest, 30 km from Bamako and the10th

of December

1998 in mount Manding in Siby, 60 km from Bamako, dried, pulverized and later

analyzed.

The leaves from Stereospermum kunthianum were analyzed. An extract was done with

ethanol and tested for activity against Candida albicans and Escherichia coli, but it did

not have any effect against these microorganisms.

Samake (2000)





51

Extraction:


/ Ethanol (80º)

________________________/ \


Residue

/ H2O 50 º C

___________________________/ \


Residue

/ H2O 100 º C

_____________________________/ \



Tests:


All solutions were hydrolyzed before testing, to separate polysaccharides into separate





different plants. Standard substance used was mannitol.


Results:



Tested

substance

Tested

part


(µg/ml)

S.k.

50ºC

Leaves + + + 0 + + + + 0 50

S.k

100ºC

Leaves + + + 0 + + + + 0 25

S.k.

50ºC

Stem

bark

+ + + 0 + + + + 0 < 15

S.k.

100ºC

Stem

bark

+ + + 0 + + + + 0 < 15

S.k. = Stereospermum kunthianum

52

Gas chromatography:


Monosaccharides Extract at 50ºC (leaves) Extract at 100ºC (leaves)

Sugar ratio (%) Test substance

ratio (%)


ratio (%)

Arabinose 7 1 7 2

Rhamnose 16 3 14 4

Xylose 2 0,3 2 0,5

Fucose 0 0 0 0

Mannose 5 1 7 2

Galactose 10 2 10 3

Glucose 53 10 38 10

Glucuronic acid 7 1 5 1

Galacturnoic

acid

0 0 17 4

Total 100 18 100 26

Polysaccharide

content (%)

18 26

Polysaccharide

content in stem

bark (%)

40 80

The stem bark showed very good effect on the complement system. This might be caused

by the high content of polysaccharides in the stem bark.

For more information about this plant see Aasberg (2001).

3.1.10 Bombacaceae

3.1.10.1 Bombax costatum Pellegr. & Vuillet


Traditional uses:

In Senegal, the stem bark and roots are known to have good diuretic properties and the

leaves are known to be rich in mucilages. The emollient properties are undisputable and

the different plant organs are used either alone or combined with other plants, especially

to treat vaginal discharge and diarrhoea (Kerharo and Adam, 1974).

This gum has traditionally been used to treat dental affections (Sanogo, 1999).

Sanogo (1999)

This author did a survey of gums found from different plants in Mali and their




53

Tests:




Water (13:3:4:3)




Results:


Coumarins, saponines, tannins , mono- and polysaccharides, mucilages, sterols and

triterpenes, leucanthocyanines, carotenoids and cardiac glycosides. The content of water

is between 7 and 8,10 %. Ashes insoluble in HCl were 1,95 %, total ash content was 3,30

% and sulphur containing ash was 3,49 % (Table 3.2).

Thin layer chromatography: matching Rf with arabinose.

Gas chromatography:

5 monosaccharides: Arabinose (45,17 %), galactose (40,92 %), glucose (7,60 %),

mannose (3,65 %) and rhamnose (2,66 %) Only 1 of these sugars was detected using thin

layer chromatography despite the large content.



Field work: Uses against dermatosis:

Masantola-region, 7.2.04

Boua Fomba uses the gum from this plant against dermatosis. The gum is carbonized and

mixed with the oil of B. parkii and the mixture is applied twice a day.

Soundjé Coulibaly and N`tiokon Traoré are using the gum from this plant to treat

dermatosis. The gum is carbonized, mixed with the oil from B. parkii and applied once

daily.

A search using www.google.com gave 395 hits while www.PubMed.com gave 0 hits

(25.02.2005).

54

3.1.11 Caesalpiniaceae

3.1.11.1 Afzelia africana Sm.


A maceration of powdered bark is used against fever and as a stimulating agent for the

gastro-intestinal system. It is also used both internally and externally against Hansen’s

disease (leprosy). The grains are sometimes considered to be toxic, but no specific

information on use has been obtained (Kerharo and Adam, 1974)

Field work:

N’Tjobougou 5.2.04

Bourama Traoré uses this recipe to treat black dermatosis; when it itches a lot the bark of

Pterocarpus erinaceus and Afzelia africana is used to make a powder. This powder is

mixed with the powder of the bark of L. velutina. One part is used for making a decoction

that is used to wash the skin, the other part is mixed with oil and applied on the

dermatosis once per day until it is completely cured.


(25.02.2005).

3.1.11.2 Burkea africana Hook.



In Senegal, the plant is used both internally and externally against dizziness and

persistent migraines. A decoction from the bark is used as anti-neuralgic remedy

(Kerharo and Adam, 1974).

Kanta (1999)





of November 1998 in the Tienfala forest, 30 km from Bamako and the 10th

of December


analyzed.

The stem bark from Burkea africana was analyzed. An extract was done with ethanol and

tested for activity against Candida albicans and Escherichia coli. The extract did not

have any effect against E. coli, but it showed effect against C. albicans at 30 mg/ml.

Activitiy: ++ (maximum activity is marked with +++).

55

At 10 mg/ml the ethanol extract of the stem bark gave 3 active products using thin layer

chromatography plates combined with the microorganism.

Samake (2000)





Extractions:


/ Ethanol (80º)

________________________/ \


Residue

/ H2O 50 º C

___________________________/ \


Residue

/ H2O 100 º C

_____________________________/ \



Tests:









Results: Thin layer chromatography and effect on the complement system:


Tested

substance

Tested

part


(µg/ml)

B.a.

50ºC

Stem

bark

+ + + 0 + + + + 0 155

B.t.

100ºC

Stem

bark

+ + + 0 + + + + 0 115

B.a. = Burkea africana

56

Gas chromatography:




Arabinose 28 27

Rhamnose 6 15

Xylose 2 5

Mannose 1 1

Galactose 10 21

Glucose 42 18

Glucuronic acid 0 0

Galacturnoic

acid

10 13

Total 99* 100

Polysaccharides

(%)

46 27


The stem bark showed activity against C. albicans as well as activity on the complement

system.

Mathisen (Mathisen 1999) discovered a high content of proanthocyanidines with

antioxidant and radical-scavenging activities in the bark from B. africana.

A search using www.google.com gave 2 hits and www.PubMed.com gave only 1 hit

(25.02.2005).

3.1.11.3 Cassia nigricans Vahl



A decoction of the leaves is used against fever and the roots are used against worms






of November 1998 in the Tienfala forest, 30 km from Bamako and the 10th

of December


analyzed.

57

The leaves from Cassia nigricans were analyzed. An extract was done with ethanol and

tested for activity against Candida albicans and Escherichia coli. The extract did not

have any effect against E. coli, but it did show good effect against C. albicans at 30

mg/ml. Activitiy: +++ (maximum activity is marked with +++).

At 10 mg/ml the ethanol extract of the leaves gave 4 active products using thin layer


Samake (2000) This author identifed 13 plants and one mushroom used in wound healing and studied




Extraction:


/ Ethanol (80º)

________________________/ \


Residue

/ H2O 50 º C

___________________________/ \


Residue

/ H2O 100 º C

_____________________________/ \


Figure 3.1.11.3.a


Tests:









58

Results:



Tested

substance

Tested

part


(µg/ml)

C.n

50ºC

Leaves + + + 0 + + + 0 + 120

C.n.

100ºC

Leaves + + + 0 + + + + + 55

C.n. = Cassia nigricans

Gas chromatography:




ratio (%)


ratio (%)

Arabinose 15 4 11 6

Rhamnose 6 2 4 2

Xylose 9 2 4 2

Mannose 5 1 10 5

Galactose 16 4 12 6

Glucose 6 2 6 3

Glucuronic acid 51 25

Galacturnoic

acid

43 12 2 1

Total 100 7 100 50

Polysaccharide

content

27 % 50 %

The leaves from the plants show very good effect against C. albicans. The decoction

from the leaves have good effect on the complement system.


3.1.11.4 Cassia sieberiana DC.

Theses: Ekoumou (2004), Fane (2002) and Sangare (2003)


In Senegal this plant is known for its purgative and diuretic properties. It is used to treat

constipation, urinary infections, intestinal parasites, Hansen’s disease (leprosy), oedemas,

sexually transmitted diseases, schistosomiasis, weakness, sterility and renal pain. The

roots are the most common part used. Twigs with leaves (maceration or decoctions) are

used to treat infantile fever and anemia (Kerharo and Adam, 1974).

59

Ekoumou (2004) This author identified 5 recipes traditionally used to treat urinary infections and cystitis.

Both single plants and combinations of plants were analyzed.

The plants were chosen from several documents on medicinal plants used in West Africa

(Kerharo and Adam, 1974 and others).

The leaves from C. sieberiana were tested alone and with fruits from Tamarindus indica

after the following recipes: 90 % C. sieberiana + 10 % T. indica and 80 % C. sieberiana

+ 20 % T. indica.

Extractions:

20 g plant material + water (200 ml)

(15 minutes at 100 ºC)

/

Decoction


20 g plant material + water (200 ml x 24 hours)

/

Water maceration


20 g plant material + ethanol (200 ml x 24 hours)

/

Ethanol extract


20 g plant material was used for all extracts, taking account of the proportions in the

recipes. All extracts were concentrated and lyophilized before use.

Tests:



combined with DPPH.

Thin layer chromatography was performed, using the following solvent system:

Butanol : acetic acid : water (60:15:25) (BAW). The plates were treated with AlCl3 and

FeCl3 and examined using UV light at 254 and 366 nm.

60

Antifungal effect:

The ethanol extracts from all plants and recipes were tested for activity against Candida

albicans. Nystatin was used as reference compound.

Antibacterial test:

The recipe with C. sieberiana combined with fruits from T. indica was tested for

antibacterial activity against Escherichia coli and Klebsiella pneumoniae.

Results:


Leaves from C. sieberiana alone:

Flavonoids, leucoanthocyans, tannins, saponines, mono- and polysaccharides, mucilages,

sterols and/or triterpenes, coumarins, anthraquinones, reducing compounds, mucilages

and cardiac glycosides. The content of water was found to be 6 % (Table 3.2).

Leaves from C. sieberiana combined with fruits from T. indica:

Flavonoids, leucoanthycyans, tannins, saponines, mono- and polysaccharides, mucilages,

and cardiac glycosides (traces), coumarins, mucilages and anthraquinons (Table 3.2).

The thin layer chromatography showed the presence of tannins and flavonoids. This

corresponds well with the results form the phytochemical tests.

The recipe with C. sieberiana combined with fruits from T. indica showed some

antibacterial activity. Doses of 100 µg gave inhibition zones of 0 mm for E. coli and 8

mm for K. pneumonia while doses of 300 µg gave inhibition zones of 7 mm and 0 mm

respectively.

No antifungal activity was observed from the ethanol extract.

All extracts showed some antioxidant activity with a mean of 2 active compounds

detected for each extract.

Fane (2002)

This author interviewed 110 healers in the Bamako district; 50 men and 60 women. Most

were between 40-70 years old. They were asked to name known toxic plants, the use of

these plants, toxic effects and possible anti-dotes.

Maiga et al (2004) did a literature research of the 19 most cited plants




but also from the literature about the plants. (See also tables 3.1.7.1.a and 3.1.7.1.b).

Biological material:


between 20-25 g.

61

- Refrigerated blood from cattle was collected at the slaughter house in Bamako

in March 2002.

- Anopheles gambiae larvae (stadium 2) were acquired from the Department de

l’epidemiologi des affections parasitaires in Bamako.


June 2002.

- Cassia siberiana roots were collected in Missabougou and in N’tjilla in

Sikasso in December 2001.

Extractions:


hours.


3x24 hours.

A maceration was made with 150 g powdered root, 1,5 l water and 200 ml honey for 24

hours.

Tests:


Toxicity:


from 500 to 5000 mg/kg. Mortality was observed up to 3 days after administration.


extracts was from 300 to 400 mg/kg. Mortality was observed up to 3 days after

administration.


Extracts were administration orally at a concentration of 37,5 mg/kg body weight for 4

weeks. After 2 weeks, 2 mice from each group were killed And their organs were



Transaminase test.

Haemolytical tests.



Results:


Alkaloids, flavonoids, tannins, saponins, coumarins, antharquinones, reducing

compounds, mono- and polysaccharides, mucilages, carotenoids, sterols and/or

triterpenes, leucanthocyanines and cardiac glycosides. The content of water was between

7,27 and 8 %. Ashes insoluble in HCl were 4,44 %, total ash content was 4,66 % and

sulphur containing ash was 8,18 %.

62

Toxicity:


LD50 intraperitoneal: no deaths observed, but the mice were shaking and had diarrhoea.

Chronic/prolonged toxicity: no deaths observed.


mice that were only given water. One of the male mice got an abcess at the right back

paw and and enlarged pancreas.

Transaminase:



3. Serum from mice (m) that were given C. siberiana

4. Serum from mice (f) that were given C. siberiana

Table 3.1.11.4.a Results from transaminase test

Serum SGOT SGPT

1 34 26

2 65 38

3 101 18

4 139 69

SGOT increased for both groups, while SGPT decrease for male mice and increased for

female mice.

Table 3.1.11.4.b Results from larvicidal test (A. gambiae)


24 hours



Decoction C.s 20 17 85

Maceration C.s. 20 14 70

Maceration C.s with

honey

20 9 45

C.s = Cassia siberiana


Table 3.1.11.4.c Results from Lombricus terrestris (L.t) test:

Extract 90000 pp Number

of L.t.

Mortality Number

of

deaths

% dead

after 72


Distilled water 5 0 0 0 0 0

Decoction C.s 5 0 0 0 0 0

Maceration C.s. 5 1 4 - 5 100

Maceration C.s

with honey

5 0 1 0 1 20

63


The plants were identified after interviews with several healers in Kendié and Finkolo

between January and February 2003. 49 healers were interviewed in Kendié and 30 in

Finkolo.

Roots from C. sieberiana were analyzed.

Tests:

Standard phytochemical tests were performed at DMT by Fane (2002).

Activity against Plasmodium falciparum (see A.1.2.1). This was tested using the extracts

from Fane; figures 3.1.11.4.a – 3.1.11.4.c

Results:



Ethanol extract: 12,30 µg/ml



The maceration with water shows effect against both A. gambiae and P. falciparum.

Although no deaths are observed, the tested extracts have some negative effects on the

tested mice.

For more information about this plant see Aasberg (2001) and Maiga et al (2004).

3.1.11.5 Cordyla pinnata (A.Rich.) Milne-Redh.



In Senegal, the stem bark and the roots are considered to be anti-helmintic due to their

purgative effects. A decoction of the leaves with salt and chili pepper is used to treat

colic. Other uses are against anorexia (weight loss), schistosomiasis, mental troubles,

aphrodisiac, diuretic and digestive and abdominal troubles (Kerharo and Adam, 1974).

This gum has traditionally been used to treat vaginal discharge (Sanogo, 1999).





64

Tests:



substances using the following solvent system: Ethyl acetate: Methanol: Acetic acid:

Water (13:3:4:3)

Gas chromatography was also performed.



Results:


Coumarins, saponines, mono- and polysaccharides, mucilages, sterols and triterpenes and

cardiac glycosides. The content of water was between 8 and 9,26 %. Ashes insoluble in

HCl were 0,65 %, total ash content was 2,42 % and sulphur containing ash was 3,35 %

(Table 3.2).


Matching Rf with galactose, galacturonic acid and glucuronic acid (they all had the same

Rf here) rhamnose and arabinose. In addition to this three spots were detected that did not

correspond with any of the references sugars.

Gas chromatography:

6 monosaccharides: Galactose (50,18 %), arabinose (24,34 %), glucuronic acid (12,01

%), mannose (6,25 %), xylose (4,13 %) and rhamnose (3,09 %). The two sugars that were

only found with gas chromatography might correspond to one or several of the

unidentified spots found from the thin layer chromatography.




(25.02.2005).

3.1.11.6 Daniellia oliveri (Rolfe) Hutch. & Dalziel

Thesis: Sanago (1999)


The most important use in Senegal is to treat colic. The resin is used either externally or

internally (or both) to treat vaginal discharge, bronchitis, renal pain, sore muscles and so

on. The roots are used as aphrodisiac by some ethnical groups, as well as against

tuberculosis. “Stipules” 10-15 cm long are used on wounds and ulcers. The roots are also

used against mental diseases comined with leaves from Ficus gnaphalocarpa (Kerharo

65

and Adam, 1974).The resin from this plant has traditionally been used to treat dental

affections (Sanogo, 1999).

Sanogo (1999)

This author did a survey of gums found from different plants in Mali and their


November 1997 and February 1998 in Bamako and Banankoro. The product obtained

from Daniellia oliveri is not a gum but a “resin” and should not contain sugars but was

still tested.

Gum1:

This gum was bought from gum salesmen so place and date of origin could not be

identified.

Gum 2:

This gum was collected in December 1997 in Banankoro.

Tests:




Water (13:3:4:3)




Results:


Gum 1:

Coumarins, saponines, tannins, mono- and polysaccharides, mucilages, sterols and

triterpenes, carotenoids and cardiac glycosides. The content of water was between 7 and


containing ash was 1,99 % (Table 3.2).

Gum 2:

Mono- and polysaccharides, mucilages, sterols and triterpenes, carotenoids and cardiac

glycosides. The content of water was between 0,85 and 1 %. Ashes insoluble in HCl were

0,93 %, total ash content was 0,41 % and sulphur containing ash was 0,45 % (Table 3.2).


Gum 1:

Matching Rf with arabinose. In addition to this there was detected one spot that did not

correspond with any of the references sugars.

Gum 2: No spots detected.

66

Results from gas chromatography:

Gum 1:

2 monosaccharides: Galactose (76,40 %) and arabinose (23,60 %). Galactose has a very

low Rf value with the solvent system used in the thin layer chromatography and might

have been detected using another solvent system.

Gum 2: No monosaccharides detected.

The product obtained form Daniellia oliveri is not a gum but a “resine” and should not

contain sugars. Gum 1 was bough from gum salesmen and has most likely been mixed

with other gums. Gum 2 was identified and collected by the author.

The tests for both Escherichia coli and Salmonella spp were negative for 1 g of both

gums. The test for starch and starch derivatives was also negative.


3.1.11.7 Piliostigma thonningii (Schumach. & Thonn.) Milne-Redh.

Thesis: Ekoumou (2004)


In Senegal, the plant is used against diarrhoea. The bark is used externally against

rheumatis as decoction or vapor bath. Leaves moisturized in hot water is rubbed on small

pox blisters too prevent scarring (Kerharo and Adam, 1974).





The stem bark from P. thonningii was tested alone and with fruits from Tamarindus

indica after the following recipes: 90 % P. thonningii + 10 % T. indica and 80 % P.

thonningii + 20 % T. indica.

Extractions:



/

Decoction


67


/

Water maceration



/

Ethanol extract



recipes.

Tests:

Standard phythochemical tests were performed at DMT.


combined with DPPH.



FeCl3 and examined using UV-light at 254 and 366 nm.

Antifungal effect: The ethanol extracts from all plants and recipes were tested for activity

against Candida albicans. Nystatin was used as reference.

Antibacterial test: The recipe with P. thonningii combined with fruits from T. indica was

tested for antibacterial activity against Escherichia coli and Klebsiella pneumoniae.

Results:


Stem bark from P. thonningii alone:

Flavonoids, leucoanthocyans, tannins, saponines, mono- and polysaccharides, mucilages,

sterols and/or triterpenes, mucilages and cardiac glycosides. The content of water was

found to be 6 % (Table 3.2).

Stem bark from P. thonningii combined with fruits from T. indica:

Flavonoids, leucoanthycyans, tannins, mono- and polysaccharides, cardiac glycosides

(traces), sterols and/or triterpenes and mucilages (Table 3.2).

The thin layer chromatography showed the presence of tannins and sterols and/or

triterpenes. This corresponds well with the results form the phytochemical tests.

68

The recipe with P. thonningii combined with fruits from T. indica showed some


mm for K. pneumonia while doses of 300 µg gave inhibition zones of respectively 7 mm

and 8 mm. This might explain the use in the treatment of urinary infections.

No antifungal activity was seen in the ethanol extract.

All extracts showed some antioxidant activity with between 1 and 3 active compounds



(25.02.2005).

3.1.12 Capparidaceae

3.1.12.1 Boscia senegalensis (Pers.) Lam. ex Poir.

Theses: Ba (1998) and Bah (1998)


In Senegal, the root, bark and leaves from this plant are said to be a very good remedy for

treating stomach ache. The plant parts are used alone or associated with roots from

Securinga virosa and/or leaves from Adansonia digitata. The plant does not have

purgative effects. The leaves are used against schistosomiasis and as a tranquilizer for

some mental diseases (Kerharo and Adam, 1974). In the Gourma district in Mali, a

decoction of dry roots is used to expel round worms in children. A maceration of fresh

leaves is used against cough. The leaves are also used to treat scabies and heart pain

(Diallo et al, 1999).

Ba (1998)

This author studied 6 plants traditionally used as “fishing poison” or used as insecticides



schistosomiasis by killing the molluscs that act as intermediate hosts.



used in all tests.

The plant material was collected the 4th

of June 1997 in Gossi. Only the leaves were

tested, and they were dried and pulverized before analysis.

Extracts were made with chloroform, methanol and water. Molluscicidal activity was

tested for all extracts at concentrations from 0,025 mg/ml to 0,4 mg/ml.

Standard phythochemical tests were performed at DMT.

69

Results:


Alkaloids (0,22 %), tannins, saponines, cardiac glycoside, coumarins, mono- and

polysaccharides, reducing compounds, mucilages and sterols and/or triterpenes. The

content of water was between 6,6 and 7,4 %. Ashes insoluble in HCl were 14,22 %, total

ash content was 6,34 % and sulphur containing ash was 6,35 % (Table 3.2).

No molluscicidal activity was observed.








The plant material was collected in Rharous in June 1996, and was dried and pulverized

before analysis. Only the leaves from B. senegalensis were analyzed.

Extractions:



_______________________/ \

DCM extract \

Residue


________________________/ \

Methanol extract \

Residue



/

Water extract

Figure 3.1.12.1.b


concentrations from 0,015 mg/ml to 0,06 mg/ml (preliminary tests) and from 1 mg/ml to

50 mg/ml later. Dead or dying larvae were registered as they floated to the surface of the

test container.

70

Results:

DCM extracts:

0,015 mg/ml and 0,03 mg/ml gave 1 % mortality while 0,06 mg/mg gave 2 % mortality.

Methanol extracts:

Only the extract at 0,06 mg/ml showed activity. It gave 1 % mortality.

Water extracts: no activity.

A review of this plant has been done by Aasberg (2001).

3.1.12.2 Maerua crassifolia Forssk.

Thesis: Bah (1998)


In Mali, a maceration of fresh leaves is mixed with butter and used against diarrhoea. A

decoction of dry leaves is used against malarial fever, jaundice and constipation. Other

uses are to cure abdominal pain, malaria fever, loss of appetite, heart disease and against

common cold (Diallo et al, 1999).









before analysis.

Only the leaves from M. crassifolia were analyzed. An extract was made with water, 500

ml x 24 hours x 3. The extracts were tested at concentrations from 0,015 mg/ml to 0,06

mg/ml on groups of 8 x 25 larvae and the number of dead larvae registered. No deaths

were observed at any of the tested concentrations.

The leaves from this plant do not show any activity in the performed tests. This plant will

not be discussed further in this thesis.


(25.02.2005).

71

3.1.13 Celastraceae

3.1.13.1 Maytenus senegalensis (Lam.) Exell

Thesis: Kanta (1999)


This plant is considered to be one of the most active in the “Pharmacopée sénégalaise”.

The main indication is a disease called “diangara Cayor”, in association with other

species. The roots are used to treat infantile diarrhoea. It is also used to treat colic, fever

and weight loss. The bark is used against fever and as a strength tonic. The leaves are

used against ocular diseases, tooth ache and mouth infections (Kerharo and Adam, 1974).




search words “vaginal discharge and vaginitis”. 18 of these plants were collected the 19th


of December


analyzed.

Leaves and stem bark from Maytenus senegalensis were analyzed. Extracts were done

with dichloromethane and methanol and tested for activity against Candida albicans and

Escherichia coli. None of the extracts had effect against these microorganisms.

The leaves and the stem bark from this plant do not show any activity in the performed

tests. This plant will not be discussed further in this thesis.


(25.02.2005).

3.1.14 Cochlospermaceae

3.1.14.1 Cochlospermum tinctorium A. Rich.



The roots are important in the treatment of children, and are used to treat rickets, colic

and helmints. The drug consist in most cases of “le tubercule vivace” which constitutes

the true medicament specific for all hepatic-biliary affections. This is believed to be

because of the yellow colored flower. The plant is also used to treat beriberi, oedemas

and tight chest (Kerharo and Adam, 1974).

72






of December


analyzed.

The roots from Cochlospermum tinctorium were analyzed. An extract was done with







Extraction:


/ Ethanol (80º)

________________________/ \


Residue

/ H2O 50 º C

___________________________/ \


Residue

/ H2O 100 º C

_____________________________/ \



Tests:









73

Results:



Tested

substance

Tested

part


(µg/ml)

C.t.

50ºC

Root

bark

+ + + 0 + + + + + 40

C.t.

100ºC

Root

bark

+ + + 0 + + + + + 45

C.t. = Cochlospermum tinctorium

Gas chromatography:




Arabinose 9 3

Rhamnose 19 6

Xylose 1 1

Mannose 2 1

Galactose 39 12

Glucose 13 70

Glucuronic acid 10 4

Galacturnoic acid 8 4

Total 101* 101*

Polysaccharides (%) 25 50


The root bark from this plant shows good activity on the complement system.

Nergård et al (Nergård 2004) studied different medicinal uses of this plant in Mali.


(25.02.2005).

3.1.15 Combretaceae

3.1.15.1 Combretum ghasalense Engl. & Diels


Sanogo (1999)

The gum from this plant has traditionally been used to treat diarrhoea. The author did a

survey of gums found from different plants in Mali and their commercial and medicinal

uses. The gums tested by this author were bought from people involved in trade/sales of

gums in November 1997. Gum number 1 is from the 1997/1998 production while gum

number 2 is from the 1996/1997 production. Both were produced in the Kolokani region

74

Tests:

Standard phythochemical tests (A.1.1.) were performed at DMT.



Water (13:3:4:3)




Results:


Gum 1:

Coumarins, mono- and polysaccharides, mucilages, sterols and triterpenes and cardiac

glycosides. The content of water was between 8 and 8,59 %. Ashes insoluble in HCl were


Gum 2:

Coumarins, mono- and polysaccharides, mucilages, sterols and triterpenes and cardiac

glycosides. The content of water was between 6 and 7,74 %. Ashes insoluble in HCl were


Gum 2:


substances and gave the following results:

Matching Rf with galactose, galacturonic acid and glucuronic acid (they all had the same

Rf here) rhamnose and arabinose. In addition to this there was detected one spot that did

not correspond with any of the references sugars. Solvent system: Ethyl acetate:

Methanol: Acetic acid: Water (13:3:4:3)


Gum 1:

5 monosaccharides: Arabinose (49,03 %), galactose (31,55 %), rhamnose (11,05 %),

galacturonic acid (5,50 %) and glucuronic acid (2,87 %). Only glucuronic acid was

undetected by the thin layer chromatography. The Rf value of glucuronic as very close to

the values of galactose and galacturonic acid.

Gum 2:

5 monosaccharides: Arabinose (53,04 %), galactose (25,63 %), rhamnose (11,08 %),

galacturonic acid (6,34 %) and glucuronic acid (2,91 %). This corresponds well with the

result from the thin layer chromatography.

The differences in the amount of the sugars found in the gum from two different seasons

vary from 5,92 % for galactose and 4,01 % for arabinose to only 0,03 % for rhamnose

and glucuronic acid.

The tests for both Escherichia coli and Salmonella spp were negative for 1 g of both

gums. The test for starch and starch derivatives was also negative.

75


3.1.15.2 Combretum glutinosum Perr.


In Senegal, this is the most prescribed drug among healers from all ethnic groups. The

leaves are used as diuretic remedy, with use in all urinary conditions. They are used to

treat hepatic-renal diseases, oedemas, yellow fever, malaria, anemia, weight loss, cough

and bronchitis. The peeled greens fruits are used to treat syphilis chancres and other

wounds. It is often used in combination with other plants to treat yellow fever,

schistosomiasis, Hansen’s disease (leprosy), weakness, spasmodic cough and also

epilepsy and mental diseases (Kerharo and Adam, 1974).

Field work:

N`Tjobougou, 5.2.04.

Djokin Diarra:

Young leaves are used against internal worms. Pound the leaves into a fine powder, mix

with porridge or liquid food like millet. Do not mix with milk. Use this treatment for 15

days.

Bourama Traoré:

Young leaves are used against otitis. Take the leaves and apply the juice directly into the

ear. This is first done one day, then investigated the next day and then used only once

more if necessary.

Konimba Traoré:

Young leaves are used against diarrhoea.

N`Pankourou Fané:

Dry leaves are pound to a fine powder. One teaspoon is mixed with hot water, filtered

and used against stomach ache. Drink as much as possible.

Djenfa Koumaré:

Trunk bark, collected from the east and west side on Thursdays, is dried and ground into

a powder. A small part is mixed in water and drunk morning against dizziness. This

treatmen is only for grown ups.

Didieni- region, 6.2.04

Diotin Traore:

Leaves are chewn against diarrhoea.

Balla Sissoko:

The Loranthus sp. is used for diagnosis. Make a decoction, take a bath and then the

healer can diagnose the patient.

76

Konimba Keita:

Young leaves are dried, powdered, mixed with water and drunk to treat dysentery.

Tietin Coulibaly:

Used against internal worms. A decoction of the leaves is drunk. Powdered leaves are

applied on external wounds.

Cho Fane:

The Loranthus sp. is used against malaria. Make a decoction and drink 3-4 times.


Sory Diarra:

A decoction of leaves is drunk against hypertension. One glass is taken per day.

Fah Diarra:

A decoction of leaves and stem bark is drunk as a tonic.

Boua Fomba:

A decoction of leaves is drunk once per day to treat leuchorrea.

Soundje Colibaly:

Decoction of young leaves is drunk against dysenteria.

N`tiokon Traore:

The powder of the Loranthus sp. is used to wash the patient if the disease is not known.

After this a diagnosis may be given.

Soungalo Traore:

Powder of trunk bark is used against wounds.

Against vaginal discharge: the young leaves are mixed with the roots of Solanum

incamon to a powder, 1 teaspoon is taken with water or porridge.


A decoction of leaves is drunk to treat constipation.

Mariba Coulibaly:

Young leaves are podwered and used against diarrhoea, the leaves may also be chewed.

Dossere Diarra:

The plant is used to treat stomach ache in small children. Make a maceration of the trunk

bark and drink this.

Naoba Diabati:

Against amenorrhoea: flowers are collected, dried and powdered. They are then mixed

with water and drunk. This will provoke bleeding.

77

Baba Diarra:

Against oedema: a decoction of the leaves is drunk morning and evening, and a bath is

also taken.

Deese Coulibaly:

A decoction of leaves is used to treat stomach ache. One teaglass or a handful is drunk.


Powder of rootbark is mixed in the food that is eaten. This is used against internal worms.

N’kodjiri Traoré:

Used as tonic: a decoction is made of the young leaves, a vapour-bath is taken and it is

drunk.

6 healers use this plant to treat stomach ache or internal worms, while 5 healers use to

treat diarrhoea or dysentery. The leaves are the plant part most commonly used.

Only one healer had no use for this plant.

Hansen (2000) studied different aspects of the structure and activity of the

polysaccharides from the bark of C. glutinosum.

Engdal (2005) and Ringerud (2003) studied different aspects of the structure and activity

of the polysaccharides from the leaves, and also in different types of leaves.


3.1.15.3 Combretum molle R. Br. ex G. Don



Inhalation of the smoke from burnt bark or branches are said to provoke aggressive

instincts in subjected persons. For this reason, the wood is never used domestically. The

smoke has been used in tribe wars to prepare for battle.

Medicinal uses are as colagogoue and against oedema. A water suspension of powdered

bark combined with gum is drunk or gargled to treat throat aches.





Tests:


78



Water (13:3:4:3)




Results:


Coumarins, mono- and polysaccharides, mucilages and cardiac glycosides. The content




Matching Rf with galactose and galacturonic (they had the same Rf here) rhamnose and

arabinose. In addition to this there was detected one spot that did not correspond with any

of the references sugars. Solvent system: Ethyl acetate: Methanol: Acetic acid: Water

(13:3:4:3)


4 monosaccharides: Galactose (39,13 %), arabinose (33,32 %), rhamnose (17,36 %) and

galacturonic acid (12,01 %). This corresponds well with the results from the thin layer

chromatography.

The tests for both Escherichia coli and Salmonella spp were negative for 1 g of gum.The

test for starch and starch derivatives was also negative.


3.1.15.4 Pteleopsis suberosa Engl. & Diels

Thesis: Ouologuem (1999)


Kerharo and Adam (1974) found no therapeutic uses of this plant, it is however

considered as “the millet medicament” and is planted together with millet if it does not

exist naturally in the area.

Ouologuem (1999)




79




Pre-made extracts were used. Water and ethanol extracts from P. suberosa twig bark was

tested.

Tests:






Results:

Larvicidal tests:


Plant extract

Time of exposure P.s. H2O P.s. Ethanol

30 minutes 0 0

1 hour 10 0

24 hours 40 45


Plant extract

Time of exposure P.s. H2O P.s. Ethanol

30 minutes 35 75

1 hour 50 80

24 hours 55 100

P.s. = Pteleopsis suberosa

The plant shows a moderate larvicidal effect. The ethanol extract has better effect than

the water extract.


(25.02.2005).

80

3.1.15.5 Terminalia avicennoides Guill et Perr.



A decoction of the roots is used to treat ascite, oedema conditions and “diangara Cayor”.

A water maceration of the roots is given to breast feeding women to treat different

diseases affecting the children they are feeding, for example conjunctivitis.

Seven leaves from the plants are used to ensure a good medicinal preparation by one

ethnic group. The root bark is used externally to treat ulcers and wounds that are hard to

heal. Powdered bark diluted in milk has emetic properties and is used by healers that are

not considered to be serious. The mixture provokes a reddish vomisement that might be

considered as the departure of the illness (Kerharo and Adam, 1974).

This plant has traditionally been used to treat haemorrhoids (Sanogo, 1999).





Tests:




Water (13:3:4:3)




Results:





(Table 3.2).


Matching Rf with galactose, arabinose and glucose (they had the same Rf here) and

rhamnose. In addition to this 2 spots were detected that did not correspond with any of

the references sugars.

81


5 monosaccharides: Arabinose (54,19 %), galactose (21,33 %), glucuronic acid (15,52

%), mannose (6,65 %) and rhamnose (2,31 %). The gas chromatography did not detect

glucose, so the spot found using thin layer chromatography marks arabinose only.

Glucuronic acid and mannose were detected in addition to the sugars found from the thin

layer chromatography. These two might correspond to one or several of the unidentified

spots found from the thin layer chromatography.

Both the types of sugars and their contents are similar to the contents found in Terminalia

macroptera (3.1.15.6).




(25.02.2005).

3.1.15.6 Terminalia macroptera Guill. et Perr.



This Terminalia specis is often confounded with the two other species: T. avicennioides

and T. laxiflora, and the uses are in many cases similar.

T. macroptera is considered an important medicament to reconstitute energy for tired or

depressed people. It is also used against fever, icterus (jaundice), syphilis and as an

aphrodisiac. The roots are considered to be diuretic and are used to treat excessive

vaginal discharge and urinary infections, especially for pregnant women.

The roots are also used to treat sterility. The leaves are used against stomach aches,

indigestion and tuberculosis. Roots and root bark is used on wounds and as eye drops

against conjunctivitis (Kerharo and Adam, 1974).

This plant has traditionally been used to treat haemorrhoids (Sanogo, 1999).




collected in February 1998 in Siby.

Tests:




Water (13:3:4:3)

82




Results:





(Table 3.2).


Matching Rf with galactose, rhamnose and arabinose. In addition to this 2 spots were

detected that did not correspond with any of the references sugars.


5 monosaccharides: Arabinose (53,8 %), galactose (25,7 %), glucuronic acid (12,4 %),

mannose (5,4 %) and rhamnose (2,7 %). Glucuronic acid and mannose were detected in

addition to the sugars found from the thin layer chromatography. These two sugars might

correspond to one or several of the unidentified spots found from the thin layer

chromatography.

Both the types of sugars and their contents are similar to the contents found in Terminalia

avicennoides.




3.1.16 Convolvulaceae

3.1.16.1 Ipomoea asarifola (Desr.) Roem. Et Schult.


In Senegal, this plant is considere to be toxic for animals, and they do not consume it.

Its main medicinal uses are gynecological. It is used to treat albuminuria for pregnant

women, bleeding outside of menstrual periods, difficult child birth and post partum

bleeding. A decotion of twigs with leaves is drunk to treat these conditions.

Other uses are in wound healing, for different eye infections and against pain in the joints


83

Field work:


Mariba Coulibaly is using the leaves and roots from this plant to treat dermatosis. Make a

decoction of the leaves and use this as a bath. Make a powder of the roots and apply on

the skin.


(25.02.2005).

3.1.17 Ebenaceae

3.1.17.1 Diospyros abyssincia Hochst. ex A. DC.


In Dioïloa in Mali this plant is used to treat stomach diseases, malaria and tooth ache. It is

also used in the healing of wounds (Odlo 2003).


The author tested the larvicidal effects of different extracts of 11 Malian plants. Larvae






Pre-made extracts were used. A methanol extract from D. abyssincia leaves was tested.

Tests:










Anopheles gambiae.

84

Results:

Larvicidal tests:


Plant extract

Time of exposure D.a. Methanol

30 minutes 30

1 hour 85

24 hours 100


Plant extract

Time of exposure D.a. Methanol

30 minutes 45

1 hour 80

24 hours 100

For the results from the extended tests at different concentrations: See tables 3.1.2.2.c -

3.1.2.2.f.

The plant shows good larvicidal effect on both tested species after 24 hours.

Odlo (2003) identified several substances with antioxidant activity from this plant.

Diallo (2000) did a survey and a phytochemial study of this plant.

3.1.18 Euphorbiaceae

3.1.18.1 Bridelia ferruginea Benth.


The author identified 29 plants used in the treatment of infections by Candida albicans,




of November 1998 in the Tienfala forest, 30 km from Bamako and 10th

December 1998

in mount Manding in Siby, 60 km from Bamako. It was dried, pulverized and later

analyzed.

Leaves, stem bark and root bark from Bridelia ferruginea were analyzed. Extracts were

done with dichloromethane and methanol and tested for activity against Candida albicans

and Escherichia coli. The extracts did not have any effect against E. coli. The

dichloromethane extracts from stem bark and root bark showed good effect against C.

albicans at 30 mg/ml. Activitiy: +++ (maximum activity is marked with +++). None of

the extracts from the leaves were active.

85

At 10 mg/ml the dichloromethane extracts of the stem bark and root bark both gave 5

active products using thin layer chromatography plates combined with the

microorganism.

The stem bark and root bark both show very good activity against C. albicans.


(25.02.2005).

3.1.18.2 Euphorbia sudanica A.Chev.

Field work:


N’Tjobougou, 5.2.04

N`Pankourou Fané uses the stem from this plant to treat black dermatosis. The stem of E.

sudanica is dried in the sun, powdered, applied on the black part of the dermatitis after

being mixed with the butter of Butyrospermum parkii. This is applied twice per day. Do

not eat this plant.


Konimba Keita uses E. sudanica to treat dermatosis. The plant is carbonized and the

powder is applied once daily.

This plant will not be discussed further in this thesis.


(25.02.2005).

3.1.18.3 Ricinus communis L.


The seeds contain ricin, which has well known purgativ and emetic properties. They are

only used to treat mental illness and leprosy.

The leaves are often used externally on wounds and blisters as antiseptic agent.

Ricin is used by one ethnic group to treat schistosomasiasis and ascite. The treatment

consists of drinking a decoction from the leaves, and a fumigation of the lower stomach


Field work:



Cho Fané uses the fruit from this plant to treat dermatosis. The fruit is carbonized, the

powder is mixed with butter of B. parkii and the mixture is applied once per day.

86


Baba Diarre uses the fruit from this plant to treat dermatosis. The fruit is carbonized, the

powder is mixed with butter of B. parkii and the mixture is applied once per day.


A review of this plant was done by Samuelsson (Samuelsson 1992).

3.1.18.4 Securinega virosa (Roxb. ex Willd.) Baill.



In Senegal, this plant is widely known and used by healers from all ethnic groups.

The most common uses are against hepatic-, biliary-, renal-, bladder- and genital trouble.

Preparations are often complicated, but are generally based on the roots.

More specific indications are: schistosomasiosis, sterility, rheumatism, fever associated

with liver disease and kidney stones.

The following plants are considered to act synergetically with S. virosa: Capparis

tomentosa, Combretum glutinosum, Combretum micranthum, Vernonia colorata,

Waltheria indica, Feretia canthioides, Momordica carantia and more (Kerharo and

Adam, 1974).

Sangare (2003)

This author tested different parts of 8 plants for activity against Plasmodium falciparum.



Finkolo.

Leaves and roots from S. virosa were collected in Blendi in the Sikasso region in May

2003.

Extractions:


/

Decoction



/

Water maceration


87



_____________ / \

Extract with DCM \

Residue


________________________/ \


Residue

/ H2O 50 º C (1 hour)

___________________________/ \


Residue

/ H2O 100 º C (1 hour)

_____________________________/ \


Figure 3.1.18.4.c Extractions with solvents of different polarity.

Tests:

Standard phytochemical tests were performed at DMT.

Activity against Plasmodium falciparum.



reagent.

Results:


Leaves:

Tannins, anthraquinones, coumarins, reducing compounds, mono- and polysaccharides

and mucilages. The content of water was between 7 and 8,03 %. Ashes insoluble in HCl

were 1,99 %, total ash content was 5,22 % and sulphur containing ash was 7,54 % (Table

3.2).

Bark:

Alkaloids (0,16 %), tannins, cardiac glycosides, sterols and/or triterpenes, coumarins,

anthraquinones, reducing compounds, mono- and polysaccharides and mucilages. The

content of water was between 7 and 7,53 %. Ashes insoluble in HCl were 1,89 %, total


The thin layer chromatography from the extract of the leaves showed one spot that

probably correspond with content of flavonoids.

88


Bark:




Leaves:







3.1.19 Fabaceae

3.1.19.1 Afrormosia laxiflora (Benth ex Baker) Harms

Theses: Kanta (1999) and Fane (2002)


In Senegal, the main uses for this plant are against pain in the joints and poisoning.

Rheumatism is treated with long baths and massages with a decoction made from the

leaves. Some forms of paralysis are treated in a similar way.

Poisioning, here in the form of bites or wounds caused by animals such as snakes, dogs

and fish, is treated both internally and externally, mostly by preparations made from the

bark, often associated with the bark from Securidaca longepedunculata.

The roots are used to treat jaundice, fever and stomach ache, and they are also said to

increase the engergy of the patient (Kerharo and Adam, 1974).

Kanta (1999) studied the root bark and Fane (2002) the leaves of this plant.

Tests:

Standard phytochemical tests were performed at DMT (Appendix A.1.1)

Extracts with dichloromethane and methanol was performed with both root bark and stem

bark from Afrormosia laxiflora. Thin layer chromatography was performed with

n-hexane – Isopropanole as solvent system. Examination of plates was performed after

treatment with Godin’s reagent and by using a UV spectrophotometer at 254 and 366 nm.

Antifungal effect was tested.

Results:

89

From the phytochemical tests.

In the leaves:

Alkaloids, flavanoids, leucoanthocyanines, tannins, saponins, cardiac glycosides, sterols

and triterpenes, coumarins, reducing compounds, mono- and polysaccharides and

mucilages (Table 3.2) (Fane, 2002).

In the root bark:

Alkaloids (0,38 %), leucoanthocyanines, tannins, saponins, cardiac glycosides, sterols

and triterpenes, carotenoids, reducing compound, mono- and polysaccharides and

mucilages (Table 3.2) (Kanta, 1999).

Antifungal effect:

30 mg/ml dichloromethane extracts of both root bark and stem bark from Afrormosia

laxiflora were active against Candida albicans.

The thin layer chromatography plates were also tested with Candida albicans and the

root bark from Afrormosia laxiflora had 7 “active spots” compared to 5 in the stem bark.








but also from the literature about the plants. (See also tables 3.1.7.1.a and 3.1.7.1.b).

Biological material:


between 20-25 g.

- Refrigerated blood from cattle was collected the slaughter house in Bamako in

March 2002.

- Anopheles gambiae larvae (stadium 2) were acquired from the Department de

l’epidemiologi des affections parasitaires in Bamako.


June 2002.

Extractions:


hours.


3x24 hours.

90

Tests:

Standard phythochemical tests (A.1.1) were performed at DMT. Results – see above.

Toxicity:


from 500 to 5000 mg/kg. Mortality was observed up to 3 days after administration.


extracts was from 300 to 400 mg/kg. Mortality was observed up tp 3 days after

administration.


Extracts were administration orally at a concentration of 28 mg/kg body weight for 4

weeks. After 2 weeks, 2 mice from each group were killed And their organs were



Transaminase test.

Haemolytical tests.



Results:

Toxicity:


LD50 intraperitoneal: no deaths observed, but the mice were shaking and had diarrhoea.

Chronic/prolonged toxicity: no deaths observed, but a decrease in weight for female

mice.


mice that were only given water.

Transaminase:



3. Serum from mice (m) that were given Afrormosia laxiflora.

4. Serum from mice (f) that were given Afrormosia laxiflora.

Table 3.1.19.1.a Results from transaminase test

Serum SGOT SGPT

1 34 26

2 65 38

3 18 96

4 186 77

SGOT increased for female mice and decreased for male mice, while SGPT increased for

both groups.

Haemostatic tests: negative

91

Table 3.1.19.1.b Results from larvicidal test on Anopheles gambiae


24 hours



Decoction A.l. 20 17 85

Maceration A.l. 20 6 30

A.l. = Afrormosia laxiflora


Table 3.1.19.1.c Results from Lombric test:

Extract

90000 pp

Number

of L.t.

Mortality Number

of deaths

% dead

after 72


Distilled

water

5 0 0 0 0 0

Decoction

A.l.

5 5 - - 5 100

Maceration

A.l.

5 0 0 0 0 0

Only the decoction displayed effect against L. terrestris.

See also Tran (2004) and Maiga et al (2004) for more information about this plant.

3.1.19.2 Bauhinia thoningii Schum.


Kanta (1999)




search words (“vaginal discharge and vaginitis”). 18 of these plants were collected the

19th


December


analyzed.

Leaves from Bauhinia thoningii were analyzed. An extract was done with ethanol,

dichloromethane and methanol and tested for activity against Candida albicans and

Escherichia coli. The extract did not have any effect against E. coli, but it did show some

effect against C. albicans at 30 mg/ml. Activitiy: ++ (maximum activity is marked with

+++). At 10 mg/ml the dichloromethane extract of the stem bark gave 2 active products

using thin layer chromatography plates combined with the microorganism.

92





Extraction:


/ Ethanol (80º)

________________________/ \


Residue

/ H2O 50 º C

___________________________/ \


Residue

/ H2O 100 º C

_____________________________/ \


Figure 3.1.19.2.a


Tests:





phthalate and methanol (1,23 : 1,66 : 100 ml) and examined using UV-light at 366w nm.


Results:



Tested

substance

Tested

part


(µg/ml)

B.t.

50ºC

Leaves + + + 0 + + + + 0 40

B.t.

100ºC

Leaves + + + 0 + + + + 0 110

B.t. = Bauhinia thoningii

93

Gas chromatography:




Arabinose 46 59

Rhamnose 24 5

Xylose 5 6

Mannose 4 2

Galactose 9 8

Glucose 4 17

Glucuronic acid 0 0

Galacturnoic

acid

12 1

Total 104* 98*

Polysaccharides

(%)

52 60


The leaves from this plant show moderate effects on the complement system. This may

be caused by the high content of polysaccharides. It demonstrates also some activity

against C. albicans.

3.1.19.3 Ostryoderris stuhlmannii (Taub.) Dunn ex. Harms.



The bark is used to treat cough, stomach ache, dysentery and as a purgative substance,

often combined with other plants. The roots are known to be emetic, and are rarely used.

An exception is the use of a maceration or decoction of the roots against fever and

internal worms by one ethnic group, only a small amount of the roots is used in this case





search words (“vaginal discharge and vaginitis”). 18 of these plants were collected the

19th


December


analyzed.

The leaves from Ostryderris sthulmannii were analyzed. An extract was done with



94





Extractions:


/ Ethanol (80º)

________________________/ \


Residue

/ H2O 50 º C

___________________________/ \


Residue

/ H2O 100 º C

_____________________________/ \



Tests:









Results:



Tested

substance

Tested

part


(µg/ml)

O.s.

50ºC

Leaves + + + 0 + + + + 0 80

O.s.

100ºC

Leaves + + + 0 + + + + 0 25

O.s. = Ostryderris sthulmannii

95

Gas chromatography:




ratio (%)


ratio (%)

Arabinose 4 1 4 1,5

Rhamnose 58 16 43 16

Xylose 1 0,3 1 0,4

Mannose 1 0,3 2 0,8

Galactose 2 0,6 19 7

Glucose 8 2,2 8 3

Glucuronic acid 26 7,84 23 9

Galacturnoic

acid

Total 100 28 100 9

Polysaccharide

content (%)

28 38

The leaves from this plant show moderate effect on the complement system.

A search using www.google.com gave 1 hit while www.PubMed.com gave 0 hits

(03.03.2005).

3.1.19.4 Prosopis africana (Guill. et Perr.) Taub.


The author identified 5 recipes traditionally used to treat urinary infections and cystitis.




The root bark from P. africana was tested alone and with fruits from Tamarindus indica

after the following recipes: 90 % P. africana + 10 % T. indica and 80 % P. africana +

20 % T. indica.

Extractions:



/

Decoction


96


/

Water maceration



/

Ethanol extract



recipes.

Tests:



combined with DPPH.




Biological tests:

Antifungal effect:

The ethanol extracts from all plants and recipes were tested for activity against Candida

albicans. Nystatin was used as reference compound.

Antibacterial test:

The recipe with P. africana combined with fruits from T. indica was tested for

antibacterial activity against Escherichia coli and Klebsiella pneumoniae.

Results:


Root bark from P. africana alone:

Alkaloids, flavonoids, anthocyanines, tannins, saponines, cardiac glycosides, mono- and

polysaccharides, mucilages, sterols and/or triterpenes, anthraquinones and reducing

compounds. The content of water was found to be 8 % (Table 3.2).

Root bark from P. africana combined with fruits from T. indica:

Alkaloids, anthocyanines, leucoanthycyans, tannins, saponines, sterols and/or triterpenes,

mono- and polysaccharides, mucilages, cardiac glucosides and mucilages (Table 3.2).

97

The thin layer chromatography showed the presence of sterols and/or triterpenes,

flavanoids and alkaloids in P. africana alone and the presence of triterpenes, tannins,

alkaloids and flavanoids in mixture of P. africana and T. indica. From the phytochemical

tests flavanoids were only detected in the pure extract, but it might be found in smaller

amounts that could only be detected by the thin layer chromatography. With this

exception the results from the thin layer chromatography corresponds well with the

results from the phytochemical tests.

The recipe with P. africana combined with fruits from T. indica showed some


mm for K. pneumonia while doses of 300 µg gave an inhibition zone of 7 mm for E. coli.

No results were observed for K. pneumoniae.


All extracts showed some antioxidant activity with between 2 and 4 active compounds



3.1.19.5 Pterocarpus erinaceus Poir.



In Senegal, this plant has multiple uses. It is used to treat dysentery either alone or in

combination with Acacia macrostachya, Lannea acida and Ostryderris stuhlmannii.

Powdered bark or roots are inhaled in the nose, or mixed with pipe tobacco to treat all

respiratory affections. A decoction of the leaves is used in inhalation, baths and beverage

to treat malaria. It is used in different combinations to treat difficult pregnancies,

weakness and yaws, and it is mainly the stem bark or roots that are used in these

preparations. The plant is also used in many medico-magical treatments (Kerharo and

Adam, 1974).

Kanta (1999)






December 1998


The stem bark from Pterocarpus erinaceus was analyzed. An extract was done with

ethanol and tested for activity against Candida albicans and Escherichia coli. The extract

98

did not have any effect against E. coli, but it showed some effect against C. albicans at 30

mg/ml. Activitiy: +, maximum activity is marked with +++.

At 10 mg/ml the ethanol extract of the stem bark gave 1 active product using thin layer






Extraction:


/ Ethanol (80º)

________________________/ \


Residue

/ H2O 50 º C

___________________________/ \


Residue

/ H2O 100 º C

_____________________________/ \



Tests:









99

Results:



Tested

substance

Tested

part


(µg/ml)

P.e.

50ºC

Stem

bark

+ + + 0 + + + + 0 < 15

P.e.

100ºC

Stem

bark

+ + + 0 + + + + 0 < 15

P.e. = Pterocarpus erinaceus

Gas chromatography:




ratio (%)


ratio (%)

Arabinose 18 1,5 13 2

Rhamnose 9 1 8 1

Xylose 11 1 4 0,5

Fucose 0 0 0 0

Mannose 3 0,2 4 0,5

Galactose 14 1 12 2

Glucose 31 2,1 40 5


Galacturnoic

acid

14 1 19 3

Total 100 7,8 100 14

Polysaccharide

content (%)

7 13

The stem bark from this plant demonstrates very good effect on the complement system.

It shows only modest activity against C. albicans.

Field work:


Bourama Toure uses this recipe to treat black dermatosis; when it itches a lot the bark of

Pterocarpus erinaceus and Afzelia africana is used to make a powder. This powder is

mixed with the powder of the bark of L. velutina. One part is used for making a decoction

that is used to wash the skin, the other part is mixed with oil and applied on the

dermatosis once per day until it is completely cured.

For a review of this plant see Aasberg (2001).

100

3.1.19.6 Pterocarpus lucens Lepr. Ex Guill. et Perr.

Kerharo and Adam (1974) reported only on use of this plant: a decoction of the roots is

said to have effect against diarrhoea.

Field work:

Dosseré Diarra uses the Loranthus sp. from this plant together with the Loranthus sp.

from Acacia ataxacanth,. Both plants are carbonized, mixed with the butter from B.

parkii and applied on affected areas.



(27.02.2005).

3.1.19.7 Stylosanthes erecta P. Beauv.



In Senegal, the plant is not widely used, however a decoction of the leaves is said to be

expectorant and a decoction of the roots is said to give energy to the patient (Kerharo and

Adam, 1974).





The aerial parts from S. erecta were tested alone and with fruits from Tamarindus indica

after the following recipes: 90 % S. erecta + 10 % T. indica and 80 % S. erecta + 20 % T.

indica (SET). S. erecta was also tested with the “calices” from Hibiscus sabdariffa using

recipes after the following recipes: 90 % S. erecta + 10 % H. sabdariffa and 80 % S.

erecta + 20 % H. sabdariffa (SEH).

Extractions:



/

Decoction


101


/

Water maceration



/

Ethanol extract



recipes.

Tests:



combined with DPPH.


Buthanol : acetic acid : water (60:15:25) (BAW). The plates were treated with AlCl3 and


Analgetic effect:

- “Writhing” test.

- “Hot-plate” test.

Haemostatic effect:

- coagulation time of blood in tests tubes was measured

- haemolytic activity was tested

Antifungal effect: The ethanol extracts from all plants and recipes were tested for activity

against Candida albicans. Nystatin was used as reference.

Antibacterial test:

The recipe with S. erecta combined with fruits from T. indica was tested for antibacterial

activity against Eshcerichia coli and Klebsiella pneumoniae.

Results:


Aerial parts from S. erecta alone:

Flavanoids, leucoanthocyans, tannins, saponines, mucilages, sterols and/or triterpenes,

coumarins, reducing compounds, mucilages and cardiac glycosides. The content of water

was found to be 6 % (Table 3.2).

102

Aerial parts from S. erecta combined with fruits from T. indica:

Leucoanthycyans, tannins, sterols and/or triterpenes, saponines, mono- and

polysaccharides, mucilages, and cardiac glycosides, coumarins and mucilages(Table 3.2).

Aerial parts from S. erecta combined with “calices” from H. sabdariffa:

Leucoanthycyans, tannins, saponines, mono- and polysaccharides, mucilages, and cardiac

glycosides, coumarins, sterols and/or triterpenes and mucilages (Table 3.2).

The thin layer chromatography of S. erecta alone showes the presence of coumarins,

flavonoids, tannins and triterpenes. This corresponds well with the results from the

phytochemical tests. The same is the situation for the SET and SEH extracts.

Antibacterial tests:

Preliminary tests:

SEH: 100 µg: no inhibition of E. coli, but 6,5 mm zone for K. pneumoniae.

300 µg: 7 mm inhibition zone of E. coli and 10 mm zone for K. pneumoniae.

STE: 100 µg: 7 mm inhibition zone of E. coli and 9 mm zone for K. pneumoniae.

300 µg: 8 mm inhibition zone of E. coli and unknown inhibition for K.

pneumoniae.

Extended tests:

Tested microbes were; E. coli (3 different strains), Staphylococcus aureus (2 strains), K.

penumoniae and Pseudomonas sp – giving a total of 8 tested bacteria types.

SEH: The ethyl acetate (400 µg/ml) and methanol (400 µg/ml) fractions were the most

active, both with activity on 5 bacteria types and with inhibition zones from 8

to14 mm.

SET: All tested extracts had good effect against E. coli and variable effects against the

other tested microbes. The DCM extracts seems to have the strongest

antimicrobial activity in total.

The alkaloid fraction was also tested: it showed good effect against E. coli, but no effect

on K. pneumoniae.


Antioxidant activity:

SET: All extracts except the water extract showed activity, with a mean of 2 active

compounds detected for each extract.

SEH: All extracts except the water extract showed activity, with between 1 and 5 active

compounds detected. The methanol extracts showed the strongest activity.

Haemostatic tests: No activity was seen.

Analgetic activity:

“Hot-plate” test: Water extract at 1000 mg/kg did not show any analgetic effect.

103

“Writhing” test:

Table 3.1.19.7 a Results of tests on analgetic activity

Tested

extract/substance

Doses ml or mg/kg Number of torsions

(M + SD)

% Protection

Distilled water 25 56,66 ± 22,01 ---

SET 20 % 500 43,33 ± 17,07 23,53

SET 20 % 1000 36,66 ± 6,43* 35,29

SEH 20 % 500 33,16 ± 10,04* 31,85

SEH 20 % 1000 28,66 ± 5,42* 49,41

Paracetamol 100 35,16 ± 9,96* 37,94

Indometacine 25 33,66 ± 3,50* 40,59

*significative results using Student t test.

The tested recipes show good antibacterial activity, especially against E. coli. They also

show analgetic activity comparable to Paracetamol and Indometacine.


(27.02.2005).

3.1.20 Flacourtiaceae

3.1.20.1 Flacourtia flabescens Forsk.







Darsalam in the Bamako district.

Pre-made extracts were used. Water and ethanol extracts from F. flabescens leaves were

tested.

Tests:






104

Results:

Larvicidal tests:


Plant extract

Time of exposure F.f. H2O F.f. Ethanol

30 minutes 0 15

1 hour 0 20

24 hours 30 80


Plant extract

Time of exposure F.f. H2O F.f. Ethanol

30 minutes 90 70

1 hour 90 90

24 hours 100 100

F.f. = Flacourtia flabescens.

F. flabescens leaves show some activity against both tested larvae species.

A search using www.google.com and www.PubMed.com gave 0 hits (27.02.2005).

3.1.21 Hypericaceae

3.1.21.1 Psorospermum senegalense Spach.

- synonym Psorospermum guineense (L.) Horch.



This plant is widely used against dermatosis all over Senegal. Internal and external

preparations from the decoction of the bark or roots are used to treat anything from mild

skin affections to syphilis, leprosy, herpes and eczema and so on.

The plant is also used to treat colics and vaginal discharge.

A fumigation of the bark is said to be used to chase demons.

The leaves are used as as expectorant, fortifier and against nausea. A decoction of the

leaves, bark and roots are used in baths and beverages to treat joint pain (Kerharo and

Adam, 1974).

Ouologuem (1999) This author tested the larvicidal effects of different extracts of 11 Malian plants. Larvae



105




Pre-made extracts were used. Water and ethanol extracts from the leaves were tested.

Tests:






Results:

Larvicidal tests:


Plant extract

Time of exposure P.g. H2O P.g. Ethanol

30 minutes 0 5

1 hour 0 5

24 hours 40 70


Plant extract

Time of exposure P.g. H2O P.g. Ethanol

30 minutes 95 5

1 hour 95 10

24 hours 95 40

P.g. = Psorospermum guineense.

P. guineense shows some larvicidal effect on the tested species.

Both names were checked:

Psorospermum senegalense


(27.02.2005).

Psorospermum guineense


(27.02.2005).

106

3.1.22 Liliaceae

3.1.22.1 Aloe buteneri

Field work - uses against dermatosis:


Konimba Traoré uses the clear succus or jelly inside the leaves of this plant directly on

dermatosis on the body. He comments that the plant is getting harder to find near his

village.


Fah Diarra uses the leaves from this plant to treat dermatosis. Cut the fresh leaves, pound

them, wash the part infected and apply the juice once per day


A search using www.google.com and www.PubMed.com gave 0 hits (27.02.2005).

3.1.23 Lycoperdeae

3.1.23.1 Podaxon aegyptiacus Mont.

Thesis: Samake (2000) The author identifed 13 plants and one mushroom used in wound healing and studied




Extraction:


/ Ethanol (80º)

________________________/ \


Residue

/ H2O 50 º C

___________________________/ \


Residue

/ H2O 100 º C

_____________________________/ \



107

Tests:









Results:



Tested

substance

Tested

part


(µg/ml)

P.a.

50ºC

Whole

plant

+ + + 0 + + + + + < 15

P.a.

100ºC

Whole

plant

+ + + + + + + + + < 15

P.a. = Podaxon aegyptiacus

Gas chromatography:




ratio (%)


ratio (%)

Arabinose 1 0,2 0 0

Rhamnose 4 0,8 5 0,75

Xylose 1 0,22 0 0

Fucose 0 0 6 1

Mannose 25 5,5 25 4

Galactose 53 12 40 6

Glucose 12 3 24 3,6

Glucuronic acid 1 0,3 0 0

Galacturnoic

acid

3 0,6 0 0

Total 100 22 100 15

Polysaccharide

content (%)

22 15

The tested extract demonstrated very good effect on the complement system.


(27.02.2005). See also Diallo et al (2002).

http://www.google.com/

108

3.1.24 Lythraceae

3.1.24.1 Lawsonia inermis L.

Theses: Ba (1998) and Ouologuem (1999)


A decoction of the leaves is used to color feet, hands and head by the women in the

Lébout and Wolof tribes. It is also know as henna. The plant is also used against

rheumatism, in difficult childbirth and abortion. It is used externally to treat sprained

muscles and/or tendons, especially in the fingers (Kerharo and Adam, 1974).

Ba (1998) This author studied 6 plants traditionally used as “fishing poison” or used as insecticides



schistosomiasis, by killing the molluscs that act as intermediate hosts.



used in all tests.

The plant material was collected in Daoudabougou the 16th

of March 1997.

Only the leaves were tested, and they were dried and pulverized before analysis.

Extraction:

Drug 150 g

/ Acetone (500 ml)

___________________ / \

Acetone extract \

Residue

/ Ethanol (500 ml)

_______________________/ \

Ethanol extract \

Residue

/ Water (500 ml)

________________________/

Water extract




0,4 mg/ml.

109

Results:


Flavanoids, tannins, cardiac glycosides, sterols and/or triterpenes, coumarins, reducing

compounds, mono- and polysaccharides and mucilages. The content of water was

between 7 and 7,05 %. Ashes insoluble in HCl were 4,32 %, total ash content was 6,62 %

and sulphur containing ash was 9,88 % (Table 3.2).


Only the acetone extracts showed activity. A concentration of 0,4 mg/ml gave 33,33 %

deaths on Biomphalaria pfeifferi and 16,66 % deaths on Bulinus truncatus.






Pre-made extracts were used. Water, ethanol, acetone and ether extracts from L. inermis

leaves were tested.

Tests:






Results:

Larvicidal test:


Plant extract

Time of exposure L.i. H2O L.i. Ethanol L.i. Acetone

30 minutes 0 5 15

1 hour 0 20 20

24 hours 15 20 55

Leaves from L. inermis show only a modest molluscicidal and larvicidal activity.


(27.02.2005).

110

3.1.25 Meliaceae

3.1.25.1 Khaya senegalensis (Desr.) A. Juss.



In Senegal, this plant is used against fever and to treat fatigue. It is also used to treat

malaria and as an abortive remedy. Decoctions or macerations of the bark is the most

common preparations used in the treatment of syphilis, vaginal discharge, leprosy and

small pox, most likely because of its purgative properties. Syphilis and leprosy are also

treated externally. This often involves use of the leaves.

The plant is used in combinations with other plants to stop vomiting and against vaginal

discharge and stomach aches (Kerharo and Adam, 1974).

The gum from the plant has traditionally been used to treat dysentery, head ache and

menstrual pain (Sanogo 1999).





Tests:




Water (13:3:4:3)





Coumarins, tannins, mono- and polysaccharides, mucilages and cardiac glycosides. The

content of water was between 8 and 8,43 %. Ashes insoluble in HCl were 3,09 %, total



Matching Rf with galactose and galacturonic acid (both had the same Rf), arabinose and

rhamnose. In addition to this there was detected one spot that did not correspond with any

of the references sugars. Solvent system: Ethyl acetate: Methanol: Acetic acid: Water

(13:3:4:3)

111


5 monosaccharides: Galactose (46,62 %), arabinose (26,79 %), galacturonic acid (13,89

%), rhamnose (10,20 %) and mannose (2,5 %). This corresponds well with the results

from the thin layer chromatography with the exception of mannose. The small content of

this sugar compared to the others might explain this difference.




3.1.25.2 Trichilia emetica Vahl.

Synonym Trichilia roka (Forssk.) Chiov.

Theses: Fane (2002), Kanta (1999) and Samake (2000)


In Senegal, the roots are known for their emetic properties and they are also used to treat

cataracts. The bark or sometimes the roots are used in the treatment of syphilis and

leprosy, often in combination with other plants. It is also used to treat poisoning, against

vaginal discharge and internal worms, as well as to treat different stomach affections;

gastritis, hepatitis, internal tumors, dyspepsia, lack of menstrual bleeding and sterility.

The plant is also used as strength tonic, stimulator of bronchial secretion and against

epilepsia. It is also believed to have magic properties by one ethnic group in Senegal


In Mali, this plant is used in the treatment of rheumatism, malaria, fever and pain. It is

also used in the treatment of chronic wounds (Diallo, 2000).




search words l (“vaginal discharge and vaginitis”). 18 of these plants were collected the

19th


of

December 1998 in mount Manding in Siby, 60 km from Bamako, dried, pulverized and

later analyzed.

The leaves from Trichilia emetica were analyzed. An extract was done with ethanol and







112

Phytochemical tests (A.1.1) were performed on 5 of these plants and, further biological

tests were performed on 3 plants: Vernonia colorata, Afrormosia laxiflora (3.1.21.1)

and Cassia siberiana (3.1.12.4). The plants were chosen from both the results from the

interviews, but also from the literature about the plants. (See also tables 3.1.7.1.a and

3.1.7.1.b). The root bark from Trichilia roka was bought at the market in Magnambougou

in July 2002.


Flavonoids, leucanthoycyans, tannins, saponins, cardiac glycoside, sterol and/or

triterpenes, coumarins, reducing compounds, mono- and polysaccharides and mucilages

(traces).The content of water was between 7,31 and 8 %. Ashes insoluble in HCl were 5,9

%, total ash content was 8,17 % and sulphur containing ash was 16,2 % (Table 3.2).





Extractions:


/ Ethanol (80º)

________________________/ \


Residue

/ H2O 50 º C

___________________________/ \


Residue

/ H2O 100 º C

_____________________________/ \



Tests:









113

Leaves from T. emetica were chosen for further tests because of their high polysaccharide

content (see results):

- Ion exchange chromatography.

- Standard phytochemical tests were performed at DMT (Appendix A.1.1.)

Results:



Tested

substance

Tested

part


(µg/ml)

T.e.

50ºC

Leaves + + + 0 + + + + + 45

T.e.

100ºC

Leaves + + + 0 + + + + + 35

T.e. = Trichilia emetica

Gas chromatography:


Monosaccharides Extract at 50ºC (leaves) Extract at 100ºC (leaves)


ratio (%)


ratio (%)

Arabinose 38 26 28 22

Rhamnose 12 8 9 8

Xylose 5 1 0,8

Fucose 0 0 0 0

Mannose 1 1 9 8

Galactose 37 26 31 26

Glucose 2 1 7 6


Galacturnoic

acid

2 2 11 9

Total 100 69 99* 82

Polysaccharide

content (%)

69 82



Alkaloids (0,1 %), flavanoids, leucoanthocyans, tannins, saponins, cardiac glycosides,

sterols and/or triterpenes, coumarins, reducing compounds, mono- and polysaccharides

and mucilages. The content of water was between 8,46 and 9 %. Ashes insoluble in HCl

were 6,55 %, total ash content was 5,6 % and sulphur containing ash was 8,645 % (Table

3.2).

Ion exchange chromatography gave a good eluation profile for the tested extract and was

the background for further separations and examination. The fractions were tested for

monosaccharide content and ICH50 in the same way as the rest of the extracts:

114

Table 3.1.25.2.c Fractions from the extract at 50ºC

Fraction % in sample Sugar ratios

Ara Rha Fuc Xyl Man Gal Glc GlcA GalA

FN 17 3 4 0 4 39 4 44 0 2

FI 50 32 10 0 1 0,2 47 0,8 6 2

FII 55 31 22 0 24 1 21 2 9 7

FIII 44 22 17 0 0,5 0,5 40 3 9 8

FIV 19 19 15 0 1 2 36 14 4 8

Table 3.1.25.2.d Fractions from the extract at 100ºC

Fraction % in

sample

Sugar ratios

Ara Rha Fuc Xyl Man Gal Glc GlcA GalA

FN 79 20 7 0 3 28 12 24 3

FI 96 42 6 0 1 2 42 2 2 3

FII 69 25 18 0 1 1 41 2 6 7

FIII 23 2 29 0 3 2 5 5 41 11

FIV 35 23 26 0 1 2 26 5 15 3

FV 18 17 2 0 2 8 40 21 7 3

FVI 14 16 8 0 7 2 29 22 14 2

The leaves from the plant have good complement activity. ICH50 for all the tested

fractions in the extended tests was < 15 µg/ml. This might explain the use of this plant in

wound healing.

Diallo (2000) performed a survey and a phytochemical study of this plant.

See also Maiga et al (2004) for a more information about the toxicity of this plant.

3.1.26 Mimosaceae

3.1.26.1 Acacia ataxacantha DC.


In Senegal, the powdered leaves are part of a combination of plants used to treat syphilis.

It is used in combination with other species to treat hernia, internal worms, wounds and

blisters. It is also used to treat gastric ulcer (Kerharo and Adam, 1974).

Field work:


Dosseré Diarra uses the Loranthus sp. from this plant together with the Loranthus sp.

from Pterocarpus lucens. Both plants are carbonized, mixed with the butter from B.

parkii and applied on affected areas.

115



(27.02.2005).

3.1.26.2 Acacia dudgeoni Craib



The plant is sometimes used against stomach aches in Senegal (Kerharo and Adam,

1974).

The gum from the plant has traditionally been used to treat ear infections (Sanogo 1999).





Tests:




Water (13:3:4:3)




Results:

The water content in the gum was between 8 and 9,6 %. The small amount of HCl -

insoluble ashes (0,64 %) corresponds well with the natural lack of sand and earth in a

gum protruding from the tree stem. Total ash content was 3,19 % and sulphur containing

ash was 4,35 % (Table 3.2). The amount of gum was too small to perform the rest of the

tests.


Matching Rf with galactose and galacturonic acid (same Rf), arabinose and rhamnose. In

addition to this there was detected two “spots” that did not correspond with any of the

references sugars. Solvent system: Ethyl acetate: Methanol: Acetic acid: Water (13:3:4:3)

Gas chromatography:

4 monosaccharides: Rhamnose (16,92 %), Glucuronic acid (19,08 %), arabinose (24,09

%) and galactose (39,91 %). This corresponds well with the results from the thin layer

116

chromatography, and it corresponds well with the sugars found in the related species

Acacia senegal and Acacia seyal.




(27.02.2005).

3.1.26.3 Acacia senegal (L.) Willd.



A decoction of the bark, or powdered gum diluted in water is used as expectorant, to calm

stomach ache and colics in both human and veterinary medicine. Gum of bad quality is

used by one of the ethnic groups in Senegal both as medicin and food, together with milk,

sugar, millet and dates.

The gum from Acacia senegal has traditionally been used to treat head aches (Sanongo,

1999).

The gum from this plant is also known as “gum arabic” and is of huge commercial

interest in the western world. It is used in different industries and the uses in food and

pharmaceutical products are the most common. Some of the uses of this gum are as an

emulsifier, tablet coating agent, stabilizer of different products and to prevent

crystallization of sugar.

Sanogo (1999) performed a survey of gums found from different plants in Mali and their



collected in the 1997 season in Kayes.

Tests:




Water (13:3:4:3)




117

Results:


Coumarins, mono- and polysaccharides, mucilages and cardiac glycosides. The content

of water was between 10 and 11,95 %. Ashes insoluble in HCl were 0,74%, total ash




addition to this there was detected one “spot” that did not correspond with any of the



4 monosaccharides: Rhamnose (14,64 %), Glucuronic acid (16,23 %), arabinose (26,32

%) and galactose (42,81 %). This corresponds well with the results from the thin layer


Acacia dudgeoni and Acacia seyal.




(27.02.2005).

3.1.26.4 Acacia seyal Del.



In Senegal, the bark is often a part of anti-leprosy treatments. A decoction is used as

purgative. The plant is also used in the treatment of jaundice and biliary fever (Kerharo

and Adam, 1974). The gum from Acacia senegal has traditionally been used to treat head

aches (Sanogo, 1999).

Sanogo (1999) did a survey of gums found from different plants in Mali and their



collected in the 1997-season in Kayes.

Tests:




Water (13:3:4:3)

118





Coumarins, tannins, mono- and polysaccharides, sterols and triterpenes, mucilages,

cardiac glycosides and leucoanthocyanines. The content of water was between 7 and 8,86

%. Ashes insoluble in HCl were 1,88%, total ash content was 3,08 % and sulphur



substances and gave the following results:


addition to this there was detected two spots that did not correspond with any of the



4 monosaccharides: Rhamnose (3,99 %), Glucuronic acid (12,68 %), arabinose (39,43 %)

and galactose (43,90 %). This corresponds well with the results from the thin layer


Acacia dudgeoni and Acacia senegal.




(27.02.2005).

3.1.26.5 Entada africana Guill. et Perr.

Theses: Ba (1998), Bah (1998) and Ouologuem (1999)


This plant is widely used in traditional medicine in different countries in West Africa.

In Mali it is used against fever (Bah, 1998), in wound healing and in the treatment of

hepatitis (Diallo et al 2000). In Senegal, the plant is used against bronchitis and cough, in

wound healing and as a magical remedy (Kerharo and Adam, 1974).





119



used in all tests.

Only the roots were tested, and they were dried and pulverized before analysis.

Extractions:

Drug 150 g

/ Dichloromethane

___________________ / \

Dichloromethane extract \

Residue

/ Methanol

_______________________/ \

Methanol extract \

Residue

/ Water

________________________/

Water extract


A liquid-liquid extraction was performed with the methanol extract:

15 g methanol extract

/

/

Water (300 ml) + Ether(300 ml)

/ \

_____________________/ \

Ether phase \

/ Water phase

Ether extract / + Butanol (300 ml)

________________________/ \

Water phase \

/ Butanol phase

Water extract \

Butanol extract

Figure 3.1.26.5.b Liquid-liquid extraction

Tests:



0,4 mg/ml.

120

Results:


Alkaloids (0,27 %), flavanoids, leucoanthocyans, tannins, saponins, cardiac glycosides,

sterols and/or triterpenes, coumarins, carotenoids, reducing compounds, mono- and


insoluble in HCl were 9,75 %, total ash content was 7,66 % and sulphur containing ash

was 7,33 % (Table 3.2).


All extracts show some activity. The strongest activity is found for the butanol extract. It

gives 100 % deaths at 0,1 mg/ml for both mollusc species.








The plant material was collected in Tchinboni in June 1996, and was dried and pulverized

before analysis.

Only the roots from E. africana were analyzed. An extract was made with water, 500 ml

x 24 hours x 3. The extracts were tested at concentrations from 0,015 mg/ml to 0,06

mg/ml on groups of 8 x 25 larvae and the number of dead larvae registered. No deaths





Larvae from Culex quinquefasciatus were collected in N’Tomikorobougou and Darsalam


Pre-made extracts were used. Water, methanol and DCM extracts from E. africana roots

were tested.

Tests:






121

Results:

Larvicidal test:


Plant extract

Time of exposure E.a. H2O E.a. Methanol E.a. DCM

30 minutes 0 5 10

1 hour 0 10 20

24 hours 20 45 55

E.a. = Entada africana.

For more information about this plant see Diallo (2000) and Aasberg (2001).

3.1.26.6 Parkia biglobosa (Jacq.) R. Br. ex G.Don


In Senegal the bark and roots are used internally, while the leaves are used externally.

Internal uses are against sterility, bronchitis, pneumonia, leprosy and sexually transmitted

diseases. It is used alone or combined with other plants. Small bits of the bark are kept in

the mouth to treat tooth ache and caries. External uses are aginst dermatosis, skin worms,

oedema and bronchitis (Kerharo and Adam, 1974).

Field work:


Djenfa Koumaré uses this plant on black dermatosis. Trunk bark is divided in two parts.

One part is used for making a decoction, the other part is carbonized. The wound is

washed with the decoction, the carbonized part is then applied, either directly as a powder

or mixed with the butter of Butyrospermum parkii. This is done every other day.



3.1.27 Moraceae

3.1.27.1 Ficus iteophylla Miq.



This plant is used for women to start delivery. A maceration of the bark is used internally

against dysentery and externally against rheumatism. A decoction of the roots is used

against tuberculosis and mental problems, more specifically against depressions and

manic periods (Kerharo and Adam, 1974).

122


These were found after interviews of healers and salesmen of medicinal plants, and a




December 1998


Leaves, stem bark and root bark from Ficus iteophylla were analyzed. Extracts were done

with dichloromethane and methanol and tested for activity against Candida albicans and

Escherichia coli. None of the extracts had effect against E. coli. The dichloromethane

extracts from the stem bark and the root bark showed effect against C. albicans at 30

mg/ml. Activities: +++ and ++ (maximum activity is marked with +++). The extracts

from the leaves showed no effect.

At 10 mg/ml the dichloromethane extract of the stem bark and root bark gave 4 and 3

active products using thin layer chromatography plates combined with the

microorganism.

The stem bark and root bark show good antifungal activity.


(27.02.2005).

3.1.28 Moringaceae

3.1.28.1 Moringa oleifera Lam.

Thesis: Chetima (2004)


In Senegal, this plant is widely used, especially in the preparation of food.

The most common medicinal uses are for children with rickets, bronchitis, fever or neural

pain. The treatmens vary from drinking of the fresh juice from the leaves, nasal

administration of powdered roots to beverages and corporal baths with macerations from

leaves, bark and roots. For adults it has the following additional indications: oedema,

rheumatism, pain in the joints, sprained muscles and so on. The gum has astringent

properties and has varied medicinal uses (Kerharo and Adam, 1974).

In Bandiagara in Mali, a decoction of the roots is used to treat pain in the bones, a

decoction of branches with leaves and roots are used against respiratory difficulties. It is

also used against fever, conjunctivitis, constipation, internal worms, stomach ache and

diarrhoea. It is also used in the preparation of food in both Mali and Niger (Chetima,

2004).

123

Chetima (2004) studied several aspects of this plant, including use in food and medicine.

The plant material was collected in the garden of DMT in Bamako in November 2002

and it was dried and pulverized before analysis. The animals used were local rabbits, both

male and female. Their average age was 2 years and their weight was between 1,5 and 2

kg.

Extractions:


(3 hours at 100 ºC)

/

Decoction


50 g plant material + water (24 hours x 3)

/

Water maceration


50 g plant material + ethanol (24 hours x 3)

/

Ethanol extract



3 hours at 50 ºC

/

Digestion

Figure 3.1.28.1.d Water at 50 ºC (digestion)

124

Residue after decoction 8 x 62,5 g = 500 g

/ Hexane

___________________ / \

Hexane extract \

Residue

/ Chloroform

_______________________/ \

Chloroform extract \

Residue

/ Methanol

________________________/ \

Methanol extract \

Residue

/ Ethanol

________________________/ \

Etanol extract \

Residue

/ H2O 50 ºC for 3 hours

___________________________/ \

Water extract 50 º C \

Residue

/ H2O 100 ºC for 3 hours

_____________________________/ \

Water extract 100 º C Residue

Figure 3.1.28.1.e Extraction using solvents of different polarity

Tests:


Thin layer chromatography was performed, using the following solvent systems:

Chloroform : methanol : water (65:35:5)

Chloroform : methanol (70:30)

Ethyl acetate : methyl ethyl ketone : formic acid : water (50:30:10:10)

Buthanol : acetic acid : water (60:15:25) (BAW)

Ligroine : ethyl acetate (1:1)



Hexane : isopropanol (5:1)

Isopropanol : water (85:15)

The thin layer plates were treated with Godins reagent, AlCl3, and naphtoresorcinol and

examined using UV light at 254 and 366 nm.

They were also tested for antioxidant acitivity using DPPH and they were finally tested

with known sugars to test monosaccharide content.

Reference sugars: arabinose, rhamnose, galactose, xylose, mannose, glucose, glucuronic

acid and galacturonic acid.

125

Gas chromatography specifications:

FID with H2 and O2 flame.

Vector gas: helium

Speed: 1,8 ml/min

Injection temperatures: 250-260ºC

Detection tempartures: 360-370ºC

Column temperature: from 140ºC to 300ºC

Injected volume: 1µl

Biological test:

Antihypercholesterol effect.

The rabbits were fed leaves from the peanut plant (Arachis hypogea). Cholesterol was

detected in these plants in preliminary tests using thin layer chromatography with

cholesterol as reference substance.

Total cholesterol and HDL-cholesterol were measured and athrogen index (AI) was

calculated (total cholesterol:HDL).

5 groups of rabbits were tested, 4 groups for one month, and one group for 45 days.

The groups received the following:

1. distilled water

2. water decoction (from the extractions with solutions of different polarity).

3. clofibrate (100 mg/kg) for a month

4. cholesterol (400 mg/kg) for two weeks, followed by the ethanol extract (80,32

mg/kg) for a month

5. cholesterol (400 mg/kg) for 6 hours, followed by the ethanol extract for one

month.

Results:


In the leaves:

Flavanoids, leucoanthocyans, tannins, cardiac glycosides, sterols and/or triterpenes,

coumarins, reducing compounds, mono- and polysaccharides and mucilages. The content


content is 7,69 % and sulphur containing ash was 11,97 % (Table 3.2).

In the residue after decoction:

Flavanoids, leucoanthocyans, tannins, cardiac glycosides, sterols and/or triterpenes,

coumarins, reducing compounds, mono- and polysaccharides and mucilages. The content



The results from the thin layer chromatography showed the presence of polyphenols and

flavonoids and coumarins. This confirms the results from the phytochemical tests.

Gas chromatography:

Time from injection to observation of tops: 35 minutes.

126

Table 3.1.28.1.a Monosaccharide composition in the water digestion (50 ºC) and

the decoction

Digestion Decoction

Sugar Quantity Ratio Quantity Ratio

Arabinose 91 13,03 93,5 15,41

Rhamnose 110 16 93,33 15,4

Xylose 32,42 4,6 30,81 5,1

Mannose - - 9,41 1,2

Galactose 200 28,7 169,56 28

Glucose 62,5 9 60 9,91

Galacturonic acid 200 28,7 150 24,8

Total 695,93 100 % 606,61 100 %

These results correspond well with the results from the thin layer chromatography.

DPPH tested on thin layer chromatography plates: All extracts showed activity except the

hexane, chloroforme and digestion extracts from the extraction with solutions of different

polarity. The other extracts had between 2 and 8 active spots.

DPPH in solution:

IC50 was calculated to be 47,7 µg/ml for the methanol extract, 113 µg/ml for the ethanol

extract, 58,8 µg/ml for the ethanol extract from the extractions with solutions of different

polarity.

Cholesterol tests:

Group 1: increase in total cholesterol and AI and a small decrease in HDL.

Group 2: small increase in total cholesterol and a small decrease in HDL.

Group 3: decrease in both total cholesterol and HDL and increased AI.

Group 4: marked increase in total cholesterol, HDL and AI after 2 weeks of cholesterol

treatment. All values decreas after 4 weeks of treatment with the ethanol extract.

Group 5. the results are unclear.

The ethanol extract seems to have good antihypercholesterol properties while the water

extract does not seem to be very active.


(27.02.2005).

3.1.29 Olacaceae

3.1.29.1 Ximenia americana L.



In Senegal, this plant is widely used. Internally, the decoction from the fresh leaves is

said to be a good non-toxic remedy against fever. Because of this it is often used to treat

127

children. It is also used against chest pain, intestinal trouble and against internal worms.

A maceration of the roots is part of a leprosy-treatment. It is also used to treat weakness

and mental illness.

Externally, the plant is used to treat pain in the head, teeth, ears and eyes. Different

preparations made from twigs with leaves, bark or roots. Powdered root combined with

Guiera senegalensis is said to diminish syphilis chancres. The fresh leaves are used as

wound dressing and haemostatic agent (Kerharo and Adam, 1974).





19th


December


analyzed.

The leaves from Ximenia americana were analyzed. An extract was done with ethanol

and tested for activity against Candida albicans and Escherichia coli. The extract did not

have any effect against E. coli, but it showed some effect against C. albicans at 30

mg/ml. Activitiy: +, maximum activity is marked with +++.

At 10 mg/ml the ethanol extract of the stem bark gave 1 active product using thin layer

chromatography plates combined with the microorganism


their monosaccharide contents and activities on the complement system. All plants were

collected in the forest near Siby in November 1998, dried and pulverized before analyzis.

Extraction:


/ Ethanol (80º)

________________________/ \


Residue

/ H2O 50 º C

___________________________/ \


Residue

/ H2O 100 º C

_____________________________/ \



128

Tests:








Activity on the complement systems was tested on sheep erythrocytes. Results:



Tested

substance

Tested

part


(µg/ml)

X.a. Leaves

50ºC

+ + + 0 + + + + + 40

X.a. Leaves

100ºC

+ + + 0 + + + + 0 15

X.a. Root

bark

50ºC

+ + 0 + + + + + < 15

X.a. Root

bark

100ºC

+ + 0 0 0 + + + 0 45

X.a. Stem

bark

50ºC

+ + + 0 + + + + + 50

X.a. Stem

bark

100ºC

+ + + 0 + + + + + < 15

X.a. = Ximenia americana

129


Leaves Root bark Stem bark

Monosaccharides Extract at

50ºC

Extract at

100ºC

Extract at

50ºC

Extract at

100ºC

Extract at

50ºC

Extract at

100ºC

Sugar

ratio (%)

Sugar

ratio (%)

Sugar

ratio (%)

Sugar

ratio (%)

Sugar

ratio (%)

Sugar

ratio (%)

Arabinose 30 28 54 36 35 27

Rhamnose 11 8 4 2 7 6

Fucose 0 0 0 4

Xylose 2 2 0 0 3 0

Mannose 4 3 0 29 6 2

Galactose 31 15 40 33 26 10

Glucose 6 8 0 0 13 27

Glucuronic acid 1 0 0 0 0 0

Galacturnoic

acid

15 36 0 0 10 23

Total 100 100 98* 100 100 99*

Polysaccharides

content

31 % 48 % 40 % 33 % 5 % 21 %


All extracts demonstrates good activity on the complement system.

Field work:


Djokin Diarra:

The eyes are washed with a decoction made from the leaves to treat conjunctivitis.

Bourama Traoré:

The roots are used against inflammation on the chin and knees. Powdered root bark is

applied directly on the inflamattion. The powder is also applied directly on inflammation

of the teeth.

Konimba Traoré:

A decoction of the leaves is used against internal inflammations. The most important case

is when the throat is swollen due to throat infection. Breathe in the vapour with a wide

open mouth.

N`Pankourou Fané:

Dried and powdered stem bark is dried is mixed with pot ash. This is applied on the

inflammation once per day.

Djenfa Koumaré:

The Loranthus sp. is used against bad spirit. Crush the Loranthus sp. and put it on the fire

in the room to remove the bad spirit.

130


Doneke Traore:

Trunk bark powder is mixed with potash and the mixture is used on inflammation.

Diotin Traore:

Power of the roots is mixed with the oil from B. parkii. This is used against

inflammation.

Balla Sissoko:

Against vaginal discharge: 1 teaspoon of powdered leaves is taken in water

Against asthma: powder made from the root is mixed with water. This is taken once per

day.

Konimba Keita:

Make a decoction of the leaves and drink one handful to treat chest pain.

Tietin Coulibaly:

Against inflammation: make a decoction of the leaves, drink a small amount and open the

mouth and inhale the vapour while boiling.

Cho Fane:

A decoction of leaves is made and the vapour is inhaled 3 times for men and 4 times for

women to treat troath inflammation


Sory Diarra:

Powder of the root bark is mixed with hot water and drunk every morning and evening

against stomach ache.

Fah Diarra:

Against conjunctivitis: wash the eyes with a decoction of the roots twice a day.

Boua Fomba:

Against dysmenorrhoea; a decoction of roots is drunk morning and evening

Against schistosomiasis: powder of the rootbark is mixed with porridge, coffee or tea and

drunk morning and evening.

Soundje Coulibaly:

A decoction of the leaves is drunk against intoxication after snakebite or rabies etc.

N`tiokon Traore:

Powder of the root is mixed with water and drunk, or a decoction can be made. This is

used as a tonic.

131

Soungalo Traore:

Against conjunctivitis: the leaves are macerated and the eyes are washed with the

maceration.


A powder made from the root is mixed with hot water and drunk in the morning before

eating to treat stomach ache.

Mariba Coulibaly:

A decoction of leaves is made and one handful is drunk and a bath is taken against

convulsions.

Dossere Diarra and Naoba Diabati:

A decoction of the leaves is used for washing the eyes to treat conjuctivitis.

Baba Diarra:

Take powder of the trunk bark and apply it on the inflammation or wound.

Deese Coulibaly:

Against schistosomiasis: a decoction is made of the trunk bark and the fruits of

Tamarindus indica. This is drunk in the morning and in the evening before going to bed.


Against conjunctivitis: a decoction of leaves is used for washing the eyes.

Against vaginal discharge: the rootbark is drunk (no method of preparation available

here).


Delirium: a decoction of leaves is drunk and used for a bath.

6 healers use this plant against conjunctivitis while 5 use it against different forms for

inflammation. Drissa Diallo believes this plant contains alkaloids. The leaves are the

most common plant part used, followed by the stem bark.


3.1.30 Opiliaceae

3.1.30.1 Opilia celtidifolia (Guill. et Perr.) Endl. ex Walp.



The roots are said to have diuretic and purgative properties. They are used to treat

leprosy. A 12 hour maceration of branches with leaves, with added salt, is used against

132

internal worms. A decoction of the branches with leaves is recommended for tooth

problems and as a stimulating agent (Kerharo and Adam, 1974).

In Dioïla and Dogonland in Mali, the most important uses of this plant are to improve

appetite, against internal worms and malaria (Rangjord, 2003).

Sangare (2003) This author tested different parts of totally 8 plants for activity against Plasmodium

falciparum. The plants were identified after interviews with several healers in Kendié and

Finkolo between January and February 2003. 49 healers were interviewed in Kendié and

30 in Finkolo.

Leaves and bark from O. celtidifolia was collected in Blendi in the Sikasso region in May

2003.

Extractions:


/

Decoction


50 g plant material + water or ethanol (80%) (500 ml x 24 hours x 2)

/

Maceration

Figure 3.1.30.1.b Maceration with water or ethanol



_____________ / \

Extract with DCM \

Residue


________________________/ \


Residue

/ H2O 50 º C (1 hour)

___________________________/ \


Residue

/ H2O 100 º C (1 hour)

_____________________________/ \



133

Tests:


Activity against Plasmodium falciparum (see A.1.2.1).



reagent.

Results:


Leaves:

Saponins, cardiac glycosides, sterols and/or triterpenes, coumarins, reducing compounds,

mono- and polysaccharides and mucilages. The content of water was between 6 and 7,02

%. Ashes insoluble in HCl were 4,19 %, total ash content was 11,8 % and sulphur


Bark:

Anthocyans, tannins, saponins, cardiac glycosides, sterols and/or triterpenes, coumarins,

reducing compounds, mono- and polysaccharides and mucilages. The content of water

was between 8 and 8,97 %. Ashes insoluble in HCl were 1,74 %, total ash content was

8,03 % and sulphur containing ash was 9,5 % (Table 3.2).

The thin layer chromatography: no results were commented


Bark:



Decoction after digestion: 9,07 µg/ml

Ethanol extract (maceration): 11,16 µg/ml


Methanol: 13,38 µg/ml

Digestion: 13,46 µg/ml

Leaves:



Ethanol extract (maceration): 10,92 µg/ml


Digestion: 12,28 µg/ml

Methanol: 14,23 µg/ml

Decoction after digestion: 15,09 µg/ml


134

Field work:


Djokin Diarra:

Leaves and roots are used against malaria. A decoction is made of the leaves. This is

drunk and also used for taking a bath. Both dry and fresh leaves are used. If dizziness

occurs in connection with malaria, use the leaves of Tamarindus indica in the same

decoction. The roots are also used to make a decoction. This is drunk and used as a bath.

About 400 ml is drunk every day. If the patient has been ill for more than a week, divide

the portion to be taken twice per day. The treatment lasts one week.

Bourama Traoré:

The leaves are used against malaria. Make a decoction and mix it with the powder of

millet, then drink it. The roots are used in a decoction against internal worms, especially

teniasis, the 5-6-meter long worm that people can get if they eat pork.

Konimba Traoré:

A decoction is made of the leaves and is used against “Calia” (a type of inflammation).

Drink it once per day.

N`Pankourou Fané:

The leaves are used against malaria. A decoction is made, and is both drunk and used for

a bath. This is done twice per day. The same preparation is used against stomach ache.

Didieni-district, 6.2.04

Doneke Traore:

A decoction of stem and leaves is used against fever. Take one bath per day and drink

about 75 ml in the morning and the evening.

Diotin Traore:

A decoction of stem and leaves is drunk and used for a bath against stomach ache.

Balla Sissoko:

Against stomach ache: a decoction of the leaves is mixed with powder of millet and

drunk.

Konimba Keita:

Against stomach ache: make a decoction of the leaves and drink this once.

Tietin Coulibaly:

Against malaria: a decoction of the stem and leaves is used for a bath and is also drunk.

Cho Fane:

Powder of the leaves is mixed with water and this is drunk in the morning and the

evening to treat stomach ache.

135


Sory Diarra:

Used against internal worms. Powder of leaves and stem is mix with milk and drunk in

the morning. This is said to be especially effective against teniasis.

Fah Diarra:

Against constipation: a decoction of leaves and stem is drunk.

Boua Fomba:

A decoction of the leaves is drunk to treat ishias and neurological pain.

Soundje Coulibaly:

A decoction of the leaves is drunk against heart failure.

N`tiokon Traore:

Against teniasis: a decoction of leaves and stem bark is drunk.

Soungalo Traore:

Against vaginal discharge: a decoction of fresh leaves is drunk.

Against haemorrhoids: macerate fresh leaves and drink the macerate.


A decoction is made of the leaves and the stem. This is drunk a little at the time against

internal worms

Dossere Diarra:

Against inflammation: a decoction of stem and leaves is used. Take a vapour bath and

drink the decoction.

Naoba Diabati:

Against schistosomiasis: fresh leaves are pound, mixed with porridge and drunk.

Baba Diarra:

A tea glass of a decoction of the leaves is drunk in the morning and the evening to treat

stomach ache.

Deese Coulibaly:

Againts pain in the nerves: a decoction is made of the leaves and the stem bark and mixed

with butter of B.parkii. The mixture is applied on the nerves.


Used as a tonic: a decoction of the leaves is drunk.


A decoction of trunk bark or roots is used to treat anxiety. One teaglass is drunk 3 times a

day, and the decoction is also used as a bath.

136

The most common part of the plant used is the leaves, followed by the stem bark. 5

healers use this plant against malaria or fever, while 8 healers use it against stomach ache

or internal worms. 2 healers do not use this plant.

Rangjord (2003) studied the structure and activity of polysaccharides from the leaves of

O. celtidifolia.


3.1.31 Oxalidaceae

3.1.31.1 Biophytum petersianum Klotzsch







December 1998


Leaves from Biophytum petersianum were analyzed. An extract was done with ethanol

and tested for activity against Candida albicans and Escherichia coli. The extract did not

have any effect against E. coli, but it did show some effect against C. albicans at 30

mg/ml. Activitiy: ++ , maximum activity is marked with +++.

At 10 mg/ml the ethanol extract of the leaves gave 3 active products using thin layer






137

Extraction:


/ Ethanol (80º)

________________________/ \


Residue

/ H2O 50 º C

___________________________/ \


Residue

/ H2O 100 º C

_____________________________/ \




Tests:









Results:



Tested

substance

Tested

part


(µg/ml)

B.p.

50ºC

Aerial

parts

+ ? + 0 0 + + + 0 < 15

B.p.

100ºC

Aerial

parts

+ + + 0 + + + + 0 < 15

B.p. = Biophytum petersianum

138

Gas chromatography:




ratio (%)


ratio (%)


Rhamnose 8 2 10 5

Xylose 9 4

Mannose 2 1

Galactose 29 7 17 8

Glucose 9 2 9 4

Galacturnoic

acid

24 5 27 12

Total 100 4 100 46

Polysaccharide

content

25 % 46%

Field work:


Djenfa Koumaré:

A decoction of the aerial part is used against stomach ache in children. This stomach ache

may be caused by internal wounds OR WORMS???.

The aerial part and salt are used for preparation of a decoction. 1 teaspoon is given

morning and evening against cough.


Tietin Coulibaly:

Against malaria: a decoction of the aerial part is drunk and used to take a bath. This is

said to be used especially for children


Sory Diarra:

Against bad spirit, mainly fever. Powder of the aerial part is mixed with water, drunk and

used for a bath. This is done once per day and is said to be used especially for children.

Fah Diarra:

Against malaria: fresh leaves are mixed with water. Wash the body with this and drink it

once a day. This is said to be used especially for babies.

N`tiokon Traore:

Againts fever: powder of the aerial part is mixed with water and the body is washed with

this.

139

Soungalo Traore:

Against malaria: powder of the aerial part is mixed with water and drunk. Another part is

mixed with the oil of B. parkii and applied on the body.


Against stomach ache: a decoction of the aerial part is drunk.

Baba Diarra:

Against malaria. 2 fingers of powder is mixed with hot water and drunk.


Against stomach ache: a decoction of the aerial part is drunk in the morning and the

evening.

3 healers use the aerial parts of the plant against stomach ache, especially for children.

4 healers use this plant to treat malaria, and two use it to treat fewer.

One of the healers uses this plant against cough.

16 of the interviewed healers did not use this plant.


(03.04.2005).

3.1.32 Papaveraceae

3.1.32.1 Argemone mexicana L.



In Senegal, this plant is known for its calming, diuretic, cholagogue and wound healing

properties. A decoction from the leaves is drunk to treat stomach aches and used in baths

to treat muscular pain. The latex is considered a good wound dressing and is also used

against dermatitis. A maceration of the roots is used to treat vaginal discharge and

hepato-biliary problems.

The plant is known to be toxic for animals, and it might cause death due to intestinal

bleeding. This is thought to be caused by the latex and the seeds. Because of this, the

plant is only used by healers (Kerharo and Adam, 1974).

Sangare (2003) This author tested different parts of totally 8 plants for activity against Plasmodium

falciparum. The plants were identified after interviews with several healers in Kendié and

Finkolo between January and February 2003. 49 healers were interviewed in Kendié and

30 in Finkolo.

Leaves from A. mexicana were collected in Blendi in the Sikasso region in May 2003.

140

Extractions:


/

Decoction



/

Water maceration




_____________ / \

Extract with DCM \

Residue


________________________/ \


Residue

/ H2O 50 º C (1 hour)

___________________________/ \


Residue

/ H2O 100 º C (1 hour)

_____________________________/ \


Figure 3.1.32.1.c Extraction with solvents of different polarity.

Tests:





reagent.

Results:


Flavanoids, alkaloids (2,7 %), tannins, cardiac glycosides, sterols and/or triterpenes,

anthraquinones, coumarins, benzoqinones, reducing compounds, mono- and

141

polysaccharides and mucilages. The content of water was between 7 and 7,78 %. Ashes

insoluble in HCl were 2,98 %, total ash content was 12,98 % and sulphur containing ash

was 12,98 % (Table 3.2).

Several spots were dected by the thin layer chromatography showed two spots, but no

conlusions are drawn.








(27.02.2005).

3.1.33 Papilionaceae

3.1.33.1 Swartzia madagascariensis Desv

Thesis: Fane (2002)

The author interviewed 110 healers in the Bamako district; 50 men and 60 women. Most



Maiga et al (2004) did a literature research of the 19 most cited plants.

Phytochemical tests (A.1.1) were performed on 5 of these plants and, further biological

tests were performed on 3 plants: Vernonia colorata, Afrormosia laxiflora (3.1.21.1)

and Cassia siberiana (3.1.12.4). The plants were chosen from both the results from the

interviews, but also from the literature about the plants. (See also tables 3.1.7.1.a and

3.1.7.1.b).

Root bark from Swartzia madagascariensis was bought at the market in Magnambougou

in July 2002.

Results:


Alkaloids (0,14 %), flavonoids, leucanthoycyans, tannins, saponins, cardiac glycoside,

sterol and/or triterpenes, coumarins, antraquinones, carotenoids, reducing compounds,

mono- and polysaccharides, mucilages and tetrahydrocannabinols.




(27.02.2005). This plant will not be discussed further in this thesis.

For more information about this plant see Maiga et al (2004).

142

3.1.34 Poaceae

3.1.34.1 Zea mays L.


In Senegal, this plant is most known for its use in preparation of food.

Mays flour mixed with roots from Securinega virosa and bark from Vernonia colorata is

used against schistosomiasis. It is used in other combinations against ascites, vaginal

discharge and leprosy (Kerharo and Adam, 1974).

Field work:


Djenfa Koumarè uses the seeds from this plant to treat black dermatosis. The seeds are

carbonized, mixed with the butter of Butyrosermum parkii and applied on the dermatosis

once per day.


A search using www.google.com gave 293000 hits and www.PubMed.com gave 12269

hits (27.02.2005).

3.1.35 Polygalaceae

3.1.35.1 Securidaca longepedunculata Fres.

Theses: Sangare (2003) and Tolo (2002)


The roots are used both internally and externally to treat snake bites. The roots have a

characteristic smell that is believed to repel snakes.The roots are also used against

intestinal parasites, and the powdered root is taken in the nose to treat migraine and

diluted in water to start child delivery. A decoction is used internally and externally to

treat bronchitis, stomach ache, leprosy, sexually transmitted diseases and fever. The

leaves are sometimes used in the same ways.

The plant is used externally to treat rheumatism, wounds and skin worms.

As this plant is known to have emetic properties, internal preparations are carefully

prepared and sometimes filtrated before administration.

The bark is used in combination with Calotropis procera as arrow poison (Kerharo and

Adam, 1974).

Sangare (2003):

This author tested different parts of 8 plants for activity against Plasmodium falciparum.



Finkolo. Roots from S. longepedunculata were analyzed.

143

Extractions:

These were made by Bagayoko (2002). The following extracts were identified from the

results in Sangares thesis:

- Petrol ether extract

- DCM extract

- Maceration I

- Decoction I

- Methanol extract

- Ethyl acetate

- Decoction II

- Maceration II

- Buthanol

Tests:

Standard phytochemical tests (A.1.1) were performed at DMT by Bagayoko (2001).

Results are not available.


Results:


Petrol ether extract: 7,34 µg/ml


Maceration I (water): 10,75 µg/ml

Decoction I: 11,05 µg/ml


Ethyl acetate: 12,07 µg/ml

Decoction II: 15,15 µg/ml

Maceration II: 16,30 µg/ml

Buthanol: 18,28 µg/ml


Tolo (2002):

Root bark from this plant has traditionally been used to treat different tooth and mouth

diseases, in an ITM in combination with Fagara zanthoxyloides and Mangifera indica.

Roots were collected in Tienfala in November 2001, they were peeled and the bark was

dried and pulverized before analyzis.

144

Extractions:


/

Decoction

The decoction was added a few drops of octanol to reduce the amount of foam coming

from the saponines.



/

Water maceration



_______________/ \ Petrol ether


Residue


_________________/ \

Extract with DCM \

Residue

/ methanol

________________________/ \


Residue

/ H2O 50 º C (1500 ml x 3 hours)

____________________________/ \


Residue

/ H2O 100 º C (1500 ml x 3 hours)

_____________________________/ \



145

Drug 250 mg

/ Decoction 10 %

/

Water extract 30 g + 200 ml water

Petrol ether (3 x 200 ml)

_____________________/ \

Petrol ether extract \

Residue

/ Ethyl acetat (3 x 200 ml)

________________________/ \

Ethyl acetat extract \

Residue

/ Buthanol (3 x 200 ml)

/ \

Water residue Buthanol extract

Figure 3.1.35.1.d Liquid-liquid extraction

Tests:


Thin layer chromatography was perfomered using the following solvent systems:

For the polar extracts: Buthanol – acetic acid – water (60:15:25)

For the organic extracts: Ligroin – ethyl acetate (2:1)

The plates were treated with Godin’s reagents and examined using UV light at 254 and

366 nm.

Antioxidant activity using DPPH combined with the thin layer plates.

Biological test (see A.1.2.1):

Antifungal activity.

Different tests on haemolytic activity (see results).

Analgetic activity – “Hot plate” and “Writhing test”

Acute anti-inflammatory activity.

Acute toxicity of both oral and intraperitoneal administration.

LD50 was found using Trevan’s curve.

Mice of the strain Oncins France Souche 1 were used in all test. They had a body weight

between 19 and 25 g.

Results:


Alkaloids (0,2 %), flavanoids, leucoanthocyans, tannins, saponins (abundant), cardiac

glycosides, sterols and/or triterpenes, coumarins, carotenoids, reducing compounds,

mono- and polysaccharides, mucilages.

146


total ash content is 4,86 % and sulphur containing ash was 5,26 % (Table 3.2).

Thin layer chromatography showed the presence of coumarins, flavonoids and saponines.

This corresponds well with the results from the phytochemical tests.

The organic extracts showed best antioxidant activity with a maximum of 4 spots in the

ethyl acetate extract, followed by 3 spots in the methanol and buthanol extracts.

Antifungal activity: one spot was detected in the water maceration and one in the petrol

ether extract.

Haemolytic test: no clear results were found from the polar extracts. From both petrol

ether extract and the DCM extract, one spot was observed. This spot seems to correspond

with the saponine spot detected from the thin layer chromatography.

Haemostatic tests:

Figure 3.1.35.1.a Results from haemostatic tests

Extracts Coagulation time for blood (minutes)

Tube Drop Tube Drop Tube Drop Tube Drop

Tested

amount

10 µl 25 µl 50 lµ 100 µl

Negative

control

10,45 17 10,45 17 10,45 17 10,45 17

Decoction 7 (33%) 9

(47%)

11 14

(17%)

12 18 13 ---

Petrol ether

(from liquid-

liquid

separation)

7,30

(30%)

10,35

(39%)

7

(33%)

--- 15 --- --- ---

Ethyl acetet 7 (33%) 10,20 7,35

(29%)

--- 10 --- 13 ---

Buthanol 10,7 12

(29%)

11,5 16

(5%)

11,30 --- 20,5 ---

Test on coagulation time of milk:

The decoction gave a positive reaction. This might be cause by the presence of tannins.

147

Toxicity:

Table 3.1.35.1.b Results from oral administration

Dose (mg/kg) Number of animals Number of dead

animals

% dead animals

750 6 0 0

1500 6 2 33,33

3000 6 5 83,33

4500 6 4 66,67

6750 6 6 100

LD50 = 538 ± 164 mg/kg. This corresponds to 2,00 ± 0,61 g/kg of the dried plant

material.

Table 3.1.35.1.c Results from peritoneal administration

Dose (mg/kg) Number of animals Number of dead

animals

% dead animals

16,65 6 0 0

25 6 1 16,67

37,5 6 2 33,33

56,25 6 3 50

70,31 6 6 100

LD50 = 56,25 ± 5,47 mg/kg. This corresponds to 209 ± 0,02 g/kg of the dried plant

material.

Tests on analgetic activity:

Table 3.1.35.1.d Results from Hot-plate tests

Tested substance Dose

(ml or

mg/kg)

Before

administration

After administration. Reaksjonstid i sekunder og

% vis økning i reaksjonstid

Time 10 min 30 min 60 min 90 min 120 min

Vann 25 7,35 ± 1,63 3,24±1,20 5,17±0,06 7,55±3,20 3,78±1,89

Securidaca

longepedunculata

1000 3,45 ± 0,61 2,61±1,47 2,77±1,16 2,89±1,35 2,11±1,04

Tramadol 5 4,17 ± 0,81 18,3±4,2

338,84 %

24,67±4,70

491,6 %

22,18±6,49

431,89 %

9,88±3,79

136,93 %

The tested extracts showed no analgetic activity compared to Tramadol. On the contrary

it seems to have worse analgetic activity then the negative control (water). The author

thinks this might be caused by the content of saponins.

148

Table 3.1.35.1.e Results from “Writhing” tests

Tested substance Dose ml or mg/kg Number of torsiong

– mean and sd.

% protection

Distilled water 25 71,17 ± 6,49 ---

S.l. extract 250 32,00 ± 8,03 55,04

S.l. extract 500 45,00 ± 6,96 36,77

S.l. extract 1000 35,00 ± 8,62 50,80

Indometacin 25 mg 5 33,67 ± 3,50 52,69

Tramadol 5 30,50 ± 6,71 57,14

S.l. = Securidaca lonepedunculata

Anti-inflammatory tests:

Table 3.1.35.1.f Immediate anti-inflammatory activity

Tested substance Dose ml or mg/kg Right foot – left foot

(mean and sd)

% inhibition

Distilled water 25 0,111 ± 0,012 ---

S.l. extract 250 0,092 ± 0,012** 16,84

S.l. extract 500 0,087 ± 0,025** 21,35

S.l. extract 1000 0,091 ± 0,011** 18,35

ASA 100 0,074 ± 0,010** 33,53

Indometacin 5 0,089 ± 0,013** 19,40

** P < 0,01 – Student t test.

Table 3.1.35.1.f Prolonged anti-inflammatory activity

Tested

substance

Dose ml or

mg/kg

1 hour after

injection

3 hours after

injection

5 hours after

injection

Distilled water 25 0,093 ± 0,134 0,111 ± 0,012 0,091 ± 0,007

S.l. 1000 0,066 ± 0,010

(28,42 %)

0,091 ± 0,011

(18,35 %)

0,078 ± 0,011

(14,60 %)

The organic extracts from the roots demonstrate the stronges activity against Plasmodium

falciparum. They also have the largest number of antioxidant substances.

The tested extracts are toxic for mice.

The anti-inflammatory effects are comparable to indometacin and seem to be dose

dependent.

For more information about this plant see Aasberg (2001)

149

3.1.36 Rhamnaceae

3.1.36.1 Zizyphus mauritiana Lam.

Theses: Ouologuem (1999) and Yansambou (2002)


The roots are used to treat sexually transmitted diseases, while the bark is used against

stomach ache. This plant is often used in combination with other plants to treat syphilis,

poisoning and indigestion.

The bark is used alone or combined with other plants to treat stomach ache, rickets,

weight loss and powdered to treat post-partum bleeding (Kerharo and Adam, 1974).






Pre-made extracts were used. Water, acetone and ether extracts from Z. mauritiana roots

were tested.

Tests:






Results:

Larvicidal test:


Plant extract

Time of exposure Z.m. H2O Z.m.Acetone Z.m. Ether

30 minutes 0 10 0

1 hour 5 10 0

24 hours 15 90 15

Z.m. = Ziziphus mauritiana.

Yansambou (2002): This plant has traditionally been used in the treatment of diabetes

Leaves from Z. mauritiana were collected in Blendio in the Sikasso region the 17th

November 2001. Root bark was bought from healers in Médine. All plant parts were

dried and pulverized before analyzis.

150

Extractions:

250 g plant material + water (2500 ml – 3 hours)

/

Decoction


250 g plant material + water (2500 ml – 3 hours at 50ºC)

/

Water digestion

Figure 3.1.36.1.b Digestion


/

Water maceration



_______________/ \Petrol ether


Residue

/ DCM (600 ml x 3 x 24 hours)

_________________/ \

Extract with DCM \

Residue

/ methanol (500 ml x 3 x 24 hours)

________________________/ \


Residue

/ ethanol 80 % (500 ml x 3 x 24 hours)

__________________________/ \


Residue

/ H2O 50 º C (1500 ml x 3 hours)

____________________________/ \


Residue

/ H2O 100 º C (1500 ml x 3 hours)

_____________________________/ \


Figure 3.1.36.1.c Extraction with solvents of different polarity.

151

Tests:



Reagents used were Godin’s, AlCl3 and Dragendorff.

Gas chromatography:

Injected volume - 1 µl.

Carrier gas – helium.

Temperature program: 140 ºC – 170 ºC – 250 ºC – 300 ºC

Hypoglycemic activity. Rabbits between 1,5 and 2,250 kg of both sexes were used.

Groups of 5 and 5 rabbits were tested.

Results:


Leaves:




total ash content is 7,53 % and sulphur containing ash was 11,67 % (Table 3.2).

Root bark:


triterpenes, coumarins, reducing compounds and mono- and polysaccharides. The content

of water was between 5,87 and 6,66 %. Ashes insoluble in HCl were 6,47 %, total ash


Thin layer chromatograpy shows spots corresponding to flavonoids, sterols and/or

triterpenes and polyphenolic compounds. This corresponds well with the results from the


Gas chromatography:

The following sugars were detected in the water extracts made from the leaves:

arabinose, rhamnose, xylose, mannose, galactose, glucose, glucuronic acid and

galacturonic acid.

The decoction from the stem bark contained: arabinose, rhamnose and glucose.

152

Hypoglycemic activity:

Different extract were tested at different concentrations and times.

Table 3.1.36.1.b % inhibition of decoction (leaves)

% inihibition

Dose / Time

(minutes)

T30 T90 T120 T180

100 mg/kg 12,01 7,6 2,97 5,2

150 mg/kg 22,04 16,36 20,06 22,31

200 mg/kg 28,74 23,75 24,54 27,44

Metformin 22,1

mg/kg

22,64 32,64 16,81 17,43

Table 3.1.36.1.c % inhibition of digestion

% inihibition

Dose / Time

(minutes)

T30 T90 T120 T180

100 mg/kg 10,11 6 - -

150 mg/kg 17,33 19,83 - -

Metformin 22,1

mg/kg

22,64 32,64 - -

Table 3.1.36.1.d % inhibition of decoction (after extraction with other solvents)

% inihibition

Dose / Time

(minutes)

T30 T90 T120 T180

100 mg/kg 26 14,30 1,32 -

150 mg/kg 32,42 19,72 23,60 18,27

200 mg/kg 32,10 34,48 26,10 29,27

Metformin 22,1

mg/kg

22,64 32,64 16,81 17,43

Table 3.1.36.1.e % inhibition of digestion (after extraction with other solvents)

% inihibition

Dose / Time

(minutes)

T30 T90 T120 T180

100 mg/kg 9,8 3,96 1,03 5,24

200 mg/kg 14,28 10,26 2,08 10,66

Metformin 22,1

mg/kg

22,64 32,64 16,81 17,43

153

Table 3.1.36.1.f % inhibition of maceration

% inihibition

Dose / Time

(minutes)

T30 T90 T120 T180

100 mg/kg 46,71 30,34 33,48 19,12

150 mg/kg 56,02 35,46 38,49 41,96

Metformin 22,1

mg/kg

22,64 32,64 16,81 17,43

Table 3.1.36.1.g % inhibition of methanol extract

% inihibition

Dose / Time

(minutes)

T30 T90 T120 T180

50 mg/kg 9,46 4,5 2,35 5,86

75 mg/kg 13,46 21,23 9,39 10,08

100 mg/kg 30,18 38,56 25,51 24,02

Metformin 22,1

mg/kg

22,64 32,64 16,81 17,43

Table 3.1.36.1.h % inhibition of ethanol extract

% inihibition

Dose / Time

(minutes)

T30 T90 T120 T180

50 mg/kg 1,76 4,73 - 1,85

75 mg/kg 32,10 40,59 28,33 33,50

100 mg/kg 51,20 45,29 34,07 43,30

Metformin 22,1

mg/kg

22,64 32,64 16,81 17,43

Table 3.1.36.1.i % inhibition of decoction (root bark)

% inihibition

Dose / Time

(minutes)

T30 T90 T120 T180

100 mg/kg - 7,58 13,80 18,68

200 mg/kg 7,9 18,04 10,61 21,32

Metformin 22,1

mg/kg

- 13,23 10,26 18,87

Positive effects on diabetes:

- Flavonoids hva antioxidant activity and can prevent atherosclerosis, a

common complications in diabetics.

- Tannins are important for wound healing

154

The water extracts contained large amount of mucliages that made the analysis difficult

to perform. These mucilages might have a role similar to Aloe spp. in wound healing, but

this has to be studied further.

Several of the extract show a good hypoglycemic activity compared to metformin. The

activity is prolonged and dose-dependent.


(27.02.2005).

3.1.36.2 Zizyphus mucronata Willd.

Thesis: Ba (1998)


In Senegal this plant is mainly used to treat diseases in the urinary tract system.

The fruits and grains are used against enuresis. A maceration done for 48 hours of

crushed fruits with grains or just of the grains is used in almost all cases of urinary

incontinence. The roots are said to be very diuretic and they are used against hematuries

(usually caused by schistosomiasis), gonococcies, oliguria and dysuria. Other rarer

indications are against Hansen’s disease, syphilis, internal worms and even some mental

diseases (Kerharo and Adam, 1974).







used in all tests.

The plant material was found in storage at DMT the 4th

of March 1997.The leaves and

“tubercules” were tested, and they were dried and pulverized before analysis.

Extraction: Drug 150 g

/ Acetone (500 ml)

___________________ / \

Acetone extract Residue

/ Ethanol (500 ml)

_______________________/ \

Ethanol extract Residue

/ Water (500 ml)

/

Water extract


155

A juice was obtained by pressing 1248 grams of young branches with leaves under a

pressure of 200 bars. 148 ml juice was obtained. It was concentrated using a rotvapor at

50 ºC.

Tests:

Standard phythochemical tests (A.1.1) were performed on the leaves at DMT.

Molluscicidal activity was tested on extracts from leaves and the juice at concentrations

between 0,025 mg/ml to 40 mg/ml.

Results:


Flavanoids, leucoanthocyans, tannins, saponins, cyanogenic glycosides, cardiac

glycosides, sterols and/or triterpenes, coumarins, reducing compounds, mono- and


insoluble in HCl were 6,16 %, total ash content is 10,14 % and sulphur containing ash

was 13,05 % (Table 3.2).


Only the acetone extract and the juice showed activity. A concentration of 0,05 mg/ml of

the acetone extract and 0,1 mg/ml of the juice gave 100 % deaths for Biomphalaria

pfeifferi, and 0,1 mg/ml of the acetone extract and 0,1 mg/ml of the juice gave 100 %

deaths for Bulinus truncatus.


(27.02.2005).

3.1.37 Rubiaceae

3.1.37.1 Canthium acutiflorum Hiern.

Thesis: Sangare (2003) This author tested different parts of 8 plants for activity against Plasmodium falciparum.



Finkolo.

Leaves and bark from C. acutiflorum was collected in Blendi in the Sikasso region in

May 2003.

156

Extractions:


/

Decoction



/

Water maceration




_____________ / \

Extract with DCM \

Residue


________________________/ \


Residue

/ H2O 50 º C (1 hour)

___________________________/ \


Residue

/ H2O 100 º C (1 hour)

_____________________________/ \



Tests:





reagent.

157

Results:


Leaves:

Alkaloids, flavanoids, anthocyans, tannins, saponins, cardiac glycosides, sterols and/or




Bark:

Flavanoids, saponins, cardiac glycosides, anthraquinones, benzoquinones, coumarins,


was between 7 and 8,54 %. Ashes insoluble in HCl were 1,5 %, total ash content was


Both extracts show several spots from the thin layer chromatography. Some of the spots

indicates the presence of flavonoids.


Bark:





Leaves:







(27.02.2005).

3.1.37.2 Crossopteryx febrifuga (Afz. ex G.don) Benth.



In Senegal, a decoction from the bark and leaves is used to relieve all types of chest pain.

A maceration of the bark is used aginst stomach ache and vaginal discharge. The bark

and leaves are also used to increase energy and strength, and the leaves are used in the

treatment of some mental diseases. All preparations made from stem- or rootbark might

be diuretic or emetic as a side effect. This is dose dependent (Kerharo and Adam, 1974).

158





19th


December


analyzed.

The fruit from Crossopteryx febrifuga was analyzed. An extract was done with ethanol

and tested for activity against Candida albicans and Escherichia coli, but it did not have

any effect against these microorganisms.

Samake (2000) Thisa author identifed 13 plants and one mushroom used in wound healing and studied




Extractions:


/ Ethanol (80º)

________________________/ \


Residue

/ H2O 50 º C

___________________________/ \


Residue

/ H2O 100 º C

_____________________________/ \

Extrac with H2O at 100 º C Residue


Tests:









159

Results:



Tested

substance

Tested

part


(µg/ml)

C.f. 50ºC Fruit + + + 0 0 + + + 0 150

C.f.

100ºC

Fruit + + + 0 + + + + 0 120

C.f. 50ºC Stem

bark

+ + + 0 0 + + + 0 135

C.f.

100ºC

Stem

bark

+ + 0 0 + + + + 0 50

C.f. = Crossopteryx febrifuga


Stem bark Fruit

Monosaccharides Extract at 50ºC Extract at

100ºC

Extract at 50ºC Extract at

100ºC

Sugar ratio (%) Sugar ratio (%) Sugar ratio (%) Sugar ratio (%)


Rhamnose 9 4 6 5

Xylose 3 0 4 2

Mannose 0 1 0 1

Galactose 16 7 11 7

Glucose 16 39 9 5


Galacturnoic

acid

41 37 61 70

Total 101* 100 99* 100

Polysaccharide

content (%)

7 29 11 15



3.1.37.3 Feretia apodanthera Del.



The plant is used to treat different urinary- and renal affections. It is also used to treat

stomach ache, and it is used in preparations in the treatment of nausea and syphilis. It is

also used as a calming agent for agitated mental conditions (Kerharo and Adam, 1974).

160




Finkolo. Leaves and bark from F. apodanthera were collected in Blendi in the Sikasso

region in May 2003.

Extractions:


/

Decoction



/

Water maceration




_____________ / \

Extract with DCM \

Residue


________________________/ \


Residue

/ H2O 50 º C (1 hour)

___________________________/ \


Residue

/ H2O 100 º C (1 hour)

_____________________________/ \



Tests:





reagent.

161

Results:


Leaves:

Tannins, saponins, cardiac glycosides, anthraquinones, coumarins, reducing compounds,

mono- and polysaccharides and mucilages. The content of water was between 4,4 and



Bark:

Flavanoids, tannins, cardiac glycosides, sterols and/or triterpenes, coumarins, reducing

compounds, mono- and polysaccharides and mucilages. The content of water was

between 5,2 and 6,57 %. Ashes insoluble in HCl were 1,5 %, total ash content was 4,17

% and sulphur containing ash was 3,93 % (Table 3.2).

The thin layer chromatography from the extract of the leaves showed several spots that

probably correspond with content of flavonoids.


Bark:





Leaves:







(27.02.2005).

3.1.37.4 Mitracarpus scaber Zucc. ex Schult. et Schult. f

Field work:


Used against dermatosis:

Djokin Diarra used the aerial parts of this plant against leaking boils. A decoction was

used to wash the dermatosis morning and evening.



(27.02.2005).

162

3.1.37.5 Mitragyna inermis (Willd.) Kuntze

Thesis: Bah (1998)


This plant is used by many different ethnic groups in Senegal. The uses vary from

treatment of stomach ache, dysentery, schistostomatiosis, fevers and syphilis to healing of

large wounds (Kerharo and Adam, 1974).

In Mali, a decoction of leaves is used against malaria. Women use the flowers for

cosmetic purposes (Diallo et al, 1999).








The plant material was collected in Ségou in June 1996, and was dried and pulverized

before analysis. The leaves and the bark from M. inermis were analyzed.

Extractions:



________________________/ \

Methanol extract \

Residue

Figure 3.1.37.5.a


/

Water extract

Figure 3.1.37.5.b

All extracts were tested on groups of 8 x 25 larvae. The extracts were tested at

concentrations from 0,015 mg/ml to 0,06 mg/ml.

Results:

Methanol extracts:

Only the 0,06 mg/ml extract showed any effect. It gave 1 % deaths.

Water extracts: No activity was observed.


163

3.1.38 Salvadoraceae

3.1.38.1 Salvadora persica L.

Thesis: Bah (1998)


In Mali, a decoction of fresh leaves is drunk as a remedy against influenza. A porridge

made from the juice from crushed leaves is used to treat cold and cough. Pieces of thin

branches are used as tooth cleaners (Diallo et al, 1999). In Senegal, the plant is especially

known for its diuretic properties. The roots are used against rheumatism. Other uses are

against Hansen’s disease (leprosy), bronchitis, couch and asthma. It is also used to clean

teeth and mouth (Kerharo and Adam, 1974).









before analysis. Only the leaves from S. persica were analyzed.

An extract was made with water, 500 ml x 24 hours x 3. The extracts were tested at

concentrations from 0,015 mg/ml to 0,06 mg/ml on groups of 8 x 25 larvae and the

number of dead larvae registered. No deaths were observed at any of the tested

concentrations.



A review of the plant has been done by Samuelsson et al (Samuelsson 1993).

3.1.39 Sapotaceae

3.1.39.1 Butyrospermum parkii (G. Don) Kotschy


In Senegal, the ”beurre du karité” has its main uses in ointments to treat sprained or stiff

muscles and rheumatism. It is also used against migraine (Kerharo and Adam, 1974).

164

Field work:


Tietin Coulibaly uses the stem from this plant combined with the stem from Combretum

molle to treat dermatosis. The dry stems are carbonized, mixed with the butter from B.

parkii and is applied on affected areas.


Baba Diarra uses Fomes fomentarius (English: german tinder, Norwegian: kjuke) that

lives on this plant to treat dermatosis. It is made into a powder, mixed with the oil from

B. parkii and applied on the skin.

The butter or oil from this plant is commonly used in preparations used on the skin.

Kjønnøy (2000) studied the structure and activites of polysaccharides from this plant.




(27.02.2005).

3.1.40 Sterculiaceae

3.1.40.1 Cola cordifolia (Cav.) R. Br.



In Senegal, this plant has several medico-magic uses. Besides this, a maceration of the

bark is used to treat bronchitis and other pulmonary affections. A maceration of the

leaves is used against leprosy. The leaves from the twigs are used in combination with

Khaya senegalensis to start child birth. The stem bark is used externally as wound

dressing (Kerharo and Adam, 1974).

For a list of different traditional uses in Mali, see Næss (Næss 2003).

Kanta (1999): This author identified 29 plants used in the treatment of infections by Candida albicans,




19th


December


analyzed.

Stem bark from Cola cordifolia was analyzed. An extract was done with ethanol and



165

Samake (2000): This author identifed 13 plants and one mushroom used in wound healing and studied




Extraction:


/ Ethanol (80º)

________________________/ \


Residue

/ H2O 50 º C

___________________________/ \


Residue

/ H2O 100 º C

_____________________________/ \



Tests:









Results:



Tested

substance

Tested

part


(µg/ml)

C.c.

50ºC

Stem

bark

+ + + 0 + + + + 0 95

C.c.

100ºC

Stem

bark

+ + + 0 0 + + + + 60

C.c. = Cola cordifolia

166

Gas chromatography:




Arabinose 16 5

Rhamnose 19 19

Fucose 1 0

Xylose 3 2

Mannose 2 0

Galactose 26 18

Glucose 12 19

Glucuronic acid 0 3

Galacturnoic

acid

20 33

Total 99* 99*

Polysaccharides

(%)

23 35


Field work:


Bourama Traoré:

The bark is used against chronic wounds. Carbonize the bark, make a powder and apply

this directly onto the wound. The dressing consists of the leaves of Combretum

micranthum under the bandage. Renew the dressing every other day.

Djenfa Koumaré:

The bark is used against hypertension. Dry the bark, pound it and mix the powder with

water. Drink this morning and evening. Use 1 soup-spoon for 4 dl water.


Diotin Traore:

A decoction of the leaves is used as a bath against very serious diseases.

Cho Fane:

Against dysmenorrhoea: a decoction of the trunk bark is drunk 4 times a day.


Soundje Coulibaly:

A decoction of the leaves is drunk to treat mental diseases.

N`tiokon Traore

Against head ache: a decoction of the stem bark is made, and a fumigation of the head

with the vapours is performed.

167

Soungalo Traore:

Nose bleeding: a fumigation of the fruitpowder is used.

Mariba Coulibaly:

Decoct of the trunk bark is drunk by women having continuous bleeding.

Dossere Diarra:

Against vascular diseases: a decoction of the trunk bark is used in a bath and is also

drunk.

Baba Diarra:

A decoction of leaves or bark is made to treat chest pain. Take a vapour-bath and also a

bath.

N’kodjiri Traoré

Againts arthritis: a decoction of trunk bark or roots is used for a bath and one teaglass is

drunk 3 times per day.

Leaves or trunk bark are used in the different preparations.

14 healers do not use this plant.

Næss (2003) studied the structure and activites from polysaccharides found in the bark of

C. cordifolia.


(03.03.2005).

3.1.41 Ulmaceae

3.1.41.1 Celtis integrifolia Lam.

Thesis: Bah (1998)


In Senegal the root is used in the treatment of mental diseases and the pulverized leaves

are used externally or per os after dilution in water to treat oedemas and several wound

types. The pulverized root is used internally to treat rheumatism, paralysis, sterility and

weakness (Kerharo et Adam, 1974).

Bah (1998): This author studied the larvicidal effects of 10 different plant extracts on Anopheles




168




The plant material was collected in Somadougou in June 1996, and was dried and

pulverized before analysis.

Only the leaves from C. integrifolia were analyzed. An extract was made with water, 500

ml x 24 hours x 3. The extracts were tested at concentrations from 0,015 mg/ml to 0,06

mg/ml on groups of 8 x 25 larvae, and the number of dead larvae registered. No deaths





(27.02.2005).

3.1.42 Vitaceae

3.1.42.1 Cissus quadrangualris L.

Theses: Ba (1998) and Ouologuem (1999)

Some traditional uses in Sengeal:

This plant is considered dangerous or toxic and its rare therapeutic actions are external.

Decoction of branches and leaves are used as bath water during fever pains and malaria.

The Indian species is used in ayurvedic medicine. (Kerharo and Adam, 1974).

Ba (1998): This author studied 6 plants traditionally used as “fishing poison” or used as insecticides






used in all tests.

The plant material was collected the 20th

June 1997 in Gossi. Only the branches were

tested, and they were dried and pulverized before analysis.

169

Extraction:

Drug 150 g

/ Chloroform 500 ml

___________________ / \

Chloroform extract \

Residue

/ Methanol 500 ml

_______________________/ \

Methanol extract \

Residue

/ Water 500 ml

________________________/

Water extract


Tests:



0,4 mg/ml.

Results:


Alkaloids (0,11 %), tannins, cardiac glycosides, sterols and/or triterpenes, coumarins,


was between 9,11 and 10 %. Ashes insoluble in HCl were 3,57 %, total ash content is



Only the chloroform extract showed some activity. A concentration of 0,4 mg/ml caused

16,66 % deaths for both mollusc species.

Ouologuem (1999): This author tested the larvicidal effects of different extracts of 11 Malian plants. Larvae





Pre-made extracts were used. Water, methanol and CHCl3 extracts from C.

quadrangularis twigs were tested.

Tests:



170








Anopheles gambiae.

Results:

Larvicidal test:


Plant extract

Time of

exposure

C.q. H2O C.q. Methanol C.q. CHCl3 C.q. Ether

30 minutes 0 10 0 95

1 hour 20 0 0 95

24 hours 35 10 30 100

C.q. = Cissus quadrangularis.

Extended tests at different concentrations: See tables 3.1.2.2.c - 3.1.2.2.f.


171

3.2 Table of phytochemical results (with explanations)

All tests are described in the appendix (A.1.1).

Positive results are shown as + to +++++. The scales vary from author to author, some

have ++++ (4) as maximum and others use +++++ (5).

Negative results are shown as 0.

Not performed tests are shown either as 0 or an empty box.

Foam index is either shown as the divider (1000/2) or the calculated result (500).

Some chemical groups are tested using multiple tests, ex. tannins and cardiac glycosides.

Some authors does not distinguish between the results from tests and might simply state

that; “this plant contains tannins”. This should be considered when the results from the

different tests are the same.

172

173

174

175

176

177

178

179

4. Discussion and conclusions

Plants that do not show any activity in the performed tests are not included in this

discussion. The same is the case for plants that are only mentioned one or two times

during the field work, especially if the uses from the field work do not seem related to the

uses described by other authors.

A total of 82 plants have been studied. The number of tests performed on each plant and

the number of plants tested varies from thesis to thesis. Because of this, a high number of

positive tests can either signify a very active plant or that many tests have been

performed on a specific plant, see each plant for details.

Table 4.a Tests performed and number of positive reactions

Test type Number of plants with positive reaction

Activity on the complement system 17

Activity against Anopheles gambiae 13

Activity against Candida albicans 11

Activity against Culex quinquefasciatus 10

Antioxidant activity 9

Activity against Plasmodium falciparum 8

Antbacterial activity (see each plant for

specifications)

5

Molluscicidal activity (see each plant for

specifications)

5

Activity against Lombricus terrestris 4

Analgesic activity – “Writhing test” 4

Antiinflammatory activity 3

Toxicicity 3

Analgesic activity – “Hot-plate test” 1

Antihypercholesterol activity 1

Antihyperglycemic activity 1

In addition to this the monosaccharide content was examined in more than 25 plants or

gums and phytochemical tests were performed on over 50 plant parts or recipes (see 3.2).

The main reason for identifying mono- and polysaccharide content and activity on the

complement system is the focus on wound healing plants and plants used against

dermatosis (Diallo et al, 2002 and others). Other properties that are useful in wound

healing are: anti-inflammatory, antimicrobial and analgesic activities.

The other tests are all important in the identification of active plants used against

common Malian diseases: malaria, schistosomiasis, sexually transmissible diseases and

diabetes.

180

Table 4.b Plants with activities from 5 of the most common tests performed

Complement A. gambiae C. albicans C. quinqufasciatius P. falciparum

A. senegalensis A. laxiflora A. laxiflora C. arborea A. mexicana

B. africana B. senegalensis A. senegalensis C. quadrangularis C. acutiflorum

B. petersianum C. arborea B. africana D. abyssinica C. sieberiana (rp)

B. thoningii C. sieberiana (rp) B. ferruginea E. africana F. apodanthera

C. cordifolia D. abyssinica B. petersianum F. flabescens O. celtidifolia

C. febrifuga F. flabescens B. thoningii L. inermis S. longepedunculata

C. nigricans G. oppositifolius C. arborea L. velutina S. mombin

C. tinctorium L. pterocarpum C. nigricans P. senegalense S. virosa

D. oliveri L. velutina F. iteophylla P. suberosa

L. velutina M. inermis P. erinaceus Z. mauritiana

O. sthulmannii P. senegalense X. americana

P. aegyptiacus P. suberosa

P. erinaceus V. colorata

S. kunthianum

S. longepedunculata

T. roka

X. americana

Interesting plants and suggestions for further work:

(These plants are chosen based on the results from the performed tests and the type and

amount of available literature).

Limeum pterocarpum

This plant showed very good activity against A. gambiae and it is also used to treat

malaria. We have not found results from any biological tests performed on this plant.

Lannea velutina

This plant demonstrates good activity against A. gambiea and C. quinquefasciatus. It has

also good effect on the complement system (ICH50 40 and 50 µg/ml). The digestion has a

very high content of polysaccharides (81 %). Identification of active substances might be

useful.

Spondias mombin

This plant shows very good activity against P. falciparum. We have not identified other

studies on this specific activity.

Borassus flabellifer 10 of the 21 interviewed healers use the carbonized flower from this plant to treat

dermatosis. An examination of the flowers chemichal content and activities might be

interesting.

181

Bombax costatum The gum from this plant was used by two of the interviewed healers in wound healing. It

has traditionally been used to treat dental affections. No biological studies have been

found.

Securinega virosa

The leaves from this plant have demonstrates good activity against Plasmodium

falciparum. We have not found any biological tests on this subject.

Stylosanthes erecta Two recipes with this plant were tested.

1. S. erecta (80 %) with T. indica (20 %)

2. S. erecta (80 %) with H. sabdariffa (20 %)

Both recipes show good antimicrobial activity as well as good antispasmodic activity

(Writhing test). They have also a high content of antioxidants. These studies are the only

ones we have found on S. erecta.

Flacourtia flabescens This plant show good activity against both A. gambiae and C. quinquefasciatus. No

literature was found on this plant using www.google.com and www.PubMed.com.

Opilia celtidifolia

The traditional use of this plant against malaria and fever is confirmed by the good

activity against P. falciparum. Further test and identification of active substance could

prove useful.

Argemone mexicana

The leaves from this plant show very good activity against P. falciparum. Further tests on

toxicity are necessary before eventual clinical tests.

Zizyphus mauritiana The root bark from this plant show very good antidiabetic activity. Further tests on

toxicity are necessary before eventual clinical tests.

Zizyphus mucronata Acetone extract from the leaves and juice made from pressing of young branches show

very good molluscicidal activity. Further tests of the juice are recommended.

The studies performed at DMT are extensive. With new laboratory facilities (2005) and

better funding the focus of the studies is moving from chemical and in vitro tests towards

clinical studies and also studies of diagnostic methods used by healers. This is important

steps in the development of safe and effective new IMTs.

http://www.google.com/

182

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186

A. Appendix

A.1 Phytochemical methods and reagents

The following phythochemical tests are described in Matière Medical (Paris et al 1976):

Alkaloids, quinones, cardiac glycosides, tannins, coumarins, sterols and/or triterpenes,

saponins, cyanogenic glycosides and anthracene glycosides.

Tests on ashes are described in Pharmacopee francaise VIII (1965).

Tests on water content are briefly described by Nordal (Nordal, 1960).

The rest of the tests and the reagents are found and translated from the examined theses.

See also Samuelsson (1999).

A.1.1 Methods

A.1.1.1 Alkaloids

Analyzed solution:

10 g plant material (powder) + 50 ml sulphuric acid (concentrated, then diluated 1/10

with distillated water). Stir and keep stirring during 24 hours in the laboratory

temperature to obtain a maceration. Filtrate and wash with water until 50 ml of filtrate is

obtained.

Characterization:

Reference substance used is strychnine.

Use 4 test tubes and put 1 ml of the filtrate into the first 2 tubes and 1 ml strychnine into

the last two tubes. Add 5 drops of Mayer-reagent in tube 1 and 3, and 5 drops of

Dragendorff reagent in tube 2 and 4.

The results are registered according to the color intensity or amount of precipitates.

A negative test = absence of alkaloides.

If the test is positive an extraction has to be performed to verify the presence of alkaloids

in the tested plant:

25 ml filtrate is put into a decantation container. The solution is made alkaline by adding

diluted ammonia (1:1) until it is possible to smell the characteristic smell (pH = 8-9). Add

a suitable amount of chloroform: alkaline solution – chloroform (1:1).

Stir without making an emulsion. Take the organic phase and dry it using sodium

anhydrate sulphate. Filtrate the solution and divide it between two containers. Evaporate

until dry using a water bath.

Add 2 ml diluted HCl (1/10) to the first container. Divide this content between two test

tubes and add the two reagents mentioned above (content of general alcaloides – Mayer

and Dragendorff).

187

Tropane alkaloids:

Add 1 ml nitric acid to the second container. Evaporate until dry using a water bath. Cool

down, then add 10 ml acetone and then one drop at the time of a 5 % solution of KOH in

alcohol (newly prepared: 0,5 g + 10 ml ethanol).

In the presence of tropane alkaloids a violet precipitation will develop.

Quantitative determination of alkaloids:

25 ml filtrate is put into a decantation container.

Make the solution alkaline by using diluted ammonia (1:1) until it is possible to smell the

characteristic smell (pH = 8-9). Perform an extraction with 50 ml chloroform. Take the

organic phase and dry on sodium anhydrate sulphate, filtrate the chloroform phase to a

weighed container. The alkaloid mass is calculated by weighing the capsule before and

after extraction.

% alkaloids in the plant material is calculated by the following formula:

% alkaloides = mass of alkaloids x 100

mass of test sample

A.1.1.2 Polyphenolic substances

Analyzed solution:

Make a 5 % infusion. Put 5 g plant material (powder) in 100 ml boiling water in a 250 ml

container. Close the colbe and let it stay for 15 minutes. Filtrate on cotton and wash with

hot water to obtain 100 ml filtrate.

Characterizations:

A.1.1.3 Flavonoides

Anthocyanines

5 ml infusion + 5 ml H2SO4, then 5 ml NH4OH.

If the coloration becomes clearer using the acid and then goes towards a blue-violet in

alkaline solution, the plant contains anthoycyanines.

Cyanidin-reaction:

5 ml infusion + 5 ml chlorohydric alcohol + some pieces of magnesium + 1 ml isoamylic

alcohol.

A pink to orange color (flavones), pink to violet (flavonones) or a red color indicates

(flavonols, flavanonols) at the layer on top of the the isoamylic alcohol is an indication of

free flavonoids.

Reduction with Mg + HCl in alcohol will reduce most aglycones from flavonoid-

glycosides to orange or red salt.

188

Leucoanthocyanines:

Do the same reaction without the magnesium pieces and heat using a water bath for a few

minutes. In the presence of leucoanthocyanines a red to dark red or violet coloration is

developed.

Cathecols give a brown to red colour.

A.1.1.4 Tannins

5 ml infusion is put into a test tube. Add 1 ml 1 % FeCl3. A greenish or blue to black

coloration is developed in the presence of tannins.

The presence of tannins is shown by adding 1 ml concentrated HCl to 5 ml infusion. Boil

for 15 minutes. A red precipitation, soluble in amylic alcohol, is developed. The

differentiation between condensed and and gallotannins is obtained from the Stiasny

reaction.

30 ml infusion + 15 ml Stiasny-reagent. Varm på vannbad ved 90 ºC i 15-30 minutter.

Development of a precipitate indicates the presence of condensed tannins. Filtrate the

solution and saturate the solution with pulverized sodium acetate. Add 1 ml 1 % FeCl3.

The development of a blue to black color indicates the presence of condensed tannins.

A.1.1.5 Saponines

Analyzed solution:

Decoction:1 g powder is put in a 250 ml colbe + 100 ml distilled water. Keep boiling for

15 minutes, filtrate and adjust to 100 ml after it has cooled down.

Characterization:

Use 10 test tubes with numbers from 1 to10. Add 1 ml decoction in tube number 1, 2 ml

in tube number 2 and so on. Adjust the volume in each tube to 10 ml with distilled water.

Shake each tube during 15 seconds (2 agitations every second). Use the thumb to close

the tubes while shaking. Leave for 15 minutes and measure the height of the foam. If it is

less than 1 cm in all the tubes the index is less than 100. The dilution where the foam is

equal to 1 cm represents the index we look for. If the height of the foam is more than 1

cm in all the tubes it will be necessary to prepare another dilation series from the

decoction and restart the process.

Foam index = 1000

The number of the tube

189

A.1.1.6 Cardiac glycosides

Analyzed solution:

1 g powder + 10 ml 60 % alcohol and 5 ml neutral 10 % Pb acetate and heat on a boiling

water bath for 10 minutes before filtrating (cotton).

Characterization:

Shake the filtrate gently with 10 ml CHCl3 without making an emulsion.

Take out the organic phase and divide it between 3 test tubes. Evaporate at a boiling

water bath until it is dry. Add 0,4 ml isopropanol to each tube. Then add the following:

Tube 1: 1 ml Baljet-reagent

Tube 2: 1 ml Kedde-reagent

Tube 3: 1 ml Raymond-Marthoud reagent

Add 4 drops of 5 % KOH in alcohol to all three tubes.

If the reaction is positive the following colors will develop:

Tube 1: Orange

Tube 2: Red to violet

Tube 3: Violet

A.1.1.7 Sterols and/or triterpenes

Analyzed solution:

Extraction.1 g powder is put into a test tube with 20 ml ether. “Sett i kork”, shake and

leave for 24 hours. Filtrate and add some ether to obtain 20 ml.

Characterization:

Liebermann-Burchards reaction:

Evaporate 10 ml of extract until it is dry and add 1 ml acetic anydrid + 1 ml CHCl3.

Divide the content between two test tubes, one of them will be used as reference.

Use a pipette to add 1-2 ml concentrated H2SO4 at the bottom of the test tubes. Do not

stir. If there is formation of a “anneau” red to brown or violet in the zone between the two

liquids and the top layer is green or violet this indicates the presence of sterols and/or

triterpenes.

Carotenoids:

Evaporate 5 ml of extract in a container (capsule) and add 2-3 drops of a saturated

solution of SbCl3 in CHCl3 or CCl4. Positive reaction is shown as the development of a

red color that changes to blue.

A.1.1.8 Coumarins

5 ml ether extract (maceration during 24 hours) is evaporated in the air.

Add 2 ml hot water. Divide the solution between 2 test tubes and add 0,5 ml 25 %

NH4OH to one of the tubes. Mix and look for fluorescence using UV at 366 nm. An

intense fluorescence in the tube with NH4OH indicates the presence of coumarins.

190

A.1.1.9 Anthraquinones and derivates

Analyzed solutions:

Chloroform extraction. 1 g drogue + 10 ml chloroform and heat for 3 minutes using a

water bath at 100 ºC. Filtrate while it is still hot and add some chloroform to obtain 10 ml

of solution.

Hydrolysate: use the powder left after the chloroform extraction and add 10 ml water and

1 ml concentrated HCl. Keep the test tube in a boiling water bath for 15 minutes. Cool

down using cold water and filtrate.

Characterizatons:

Free anthracenes:

1 ml chloroform extract + 1 ml diluted NH4OH and stir. A red or redish color indicates

the presence of free anthraquinones.

O-glycosides:

5 ml hydrolysat + 5 ml CHCl3 is stirred. Take out the organic phase and put it into a test

tube, add 1 ml diluted NH4OH and stir well. The presence of anthraquinones is indicated

by a red color of variable intensity. If the reaction is negative or weakly positive look for

O-glycosides “a génine reduite”.

5 ml hydrolysat + 3-4 drops of 10 % FeCl3 and heat on a water bath for 5 minutes.

Cool down and shake with 5 ml CHCl3. Take out the organic phase and put it into a test

tube, add 1 ml diluted NH4OH and stir well. The presence of oxydated products of

anthroles and anthrones is indicated by a red color of variable intensity.

C-glycosides:

Use the water phase (preserved with 10 ml water) and add 1 ml 10 % FeCl3. Keep the test

tube in a boiling water bath for 30 minutes, cool down and shake with 5 ml CHCl3. Take

out the organic phase and put it into a test tube, add 1 ml diluted NH4OH and stir well.

The presence of C-glycosides is indicated by a red color of variable intensity.

Brissermoret and Combes reaction (differentiation of quinines):

1 g powder is put in a 250 ml ecolbe. Moisturise with 10 % H2SO4. Add 20 ml of a

mixture of ether and chloroform (1:1) mix and leave to make a masseration for 24 hours.

Filtrate and let evaporate in the air 5 ml of the filtrate. Add a few drops of 95 % alcohol.

Add one drop at the time of 5 % nickel acetate. According to the quinine content the

following results may be observed:

- benzoquinones: blue color and precipitate

- naphtoquinones: violet color and precipitate

- anthraquinones: red color without precipitate

A.1.1.10 Cyanogenic glycosides

1 g powder + 5 ml mixture of water and toluene (1:1).

Mix well and clean the upper of the tube. A “picrosodé” paper is connected to the upper

part of the tube using a closing cap and without the paper touching the solution.

191

A red coloration of the paper (can appear slowly or fast) indicates the presence of

cyanogenic glycosides.

A.1.1.11 Reducing compounds

5 ml 10 % water decoction is put into a suitable container (capsule) and is evaporated

until it is dry on a water bath. Add 1 ml Fehlings-reagent. A brick red precipitate is an

indication of the presence of reducing compounds.

A.1.1.12 Mono- and polysaccharides

5 ml 10 % water decoction is put into a suitable container (capsule) and is evaporated

until it is dry on a water bath. Add 2-3 drops of concentrated H2SO4. Add 3-4 drops of

alcohol saturated with thymole after 5 minutes. The development of a red coloration

indicates the presence of mono- and polysaccharides.

A.1.1.13 Mucilages

1 ml 10 % water decoction + 5 ml absolute alcohol.

The development of a non-solid precipitate indicates the presence of mucilages.

A.1.1.14 Tetrahydrocannabinols

0,5 g powder + 5 ml petrol ether. Shake for 15 minutes, filtrate into a suitable container

(capsule) and evaporate until it is dry using a water bath. Add 3-4 drops of 5 % KOH in

alcohol. A violet color indicates the presence of tetrahydrocannabinols.

A.1.1.15 Water extractible substances

This measurement gives an evaluation of how water soluble the drug is in water.

1 g of powdered plant material is put in a colbe with 20 ml of water and boiled for 15

minutes. After it is cooled down (about 20 minutes) the solution is filtrated through

cotton wool in a previously weighed container. After drying, the container is measured

and the amount of water extractible substances is calculated.

A.1.1.16 Water content

Gravimetrical contents:

Determination of weight loss of a know quantity of powder after dessication in a

temperature regulated heating chamber at 100 °C ± 3 during 24 hours.

Practical performance:

5 samples are put into 5 previously weighed suitable containers. The containers are

placed in the heating chamber for 100 °C ± 3 during 24 hours.

192

The samples are cooled down in a dessicator and measured with their containers. The

mean weight loss is calculated and converted to loss per 100 g of the tested plant.

Volumetric method:

Measurement of the amount of distilled water collected from distilling a known amount

of the tested plant in a non-water miscible solvent (toluene) at a constant temperature.

Practical performance – see one of the theses.

A.1.1.17 Tests involving ashes

These tests are performed to measure the content of foreign elements like sand and earth

in the plant tissue.

Total amount of ash:

Measurement of the total amount of ash from a certain plant is found by weighing the

amount of white ashes obtained from incinerating the plant material in an oven “moufle”

at 800 °C for 4 hours. 5 samples are tested and the mean weight is calculated. This mass

is then compared to 100 g of powdered plant.

Determination of sulfur content in ash:

This is a method used to find inorganic substances in plant material. The sample is

moistured with sulphuric acid and put in an oven at 110°C for 24 hours, then at 800°C for

4 hours. The ash that is left contains non volatile sulfates. The obtained mass is compared

to 100 g of powdered plant.

Determination of amount of ash that is not soluble in hydrochloric acid:

The total amount of ash is boiled in diluted hydrochloric acid. The insoluble matter is

collected on a filter paper.

A know amount of ash from a known amount of plant is boiled in 20 ml 10 %

hydrochloric acid for 15 minutes, after cooling it down, the solution is filtrated, washed

and heated at 110°C for 24 hours. After drying the residue is cooled down and put in an

oven at 800°C for 4 hours.

The obtained amount of ash that is not soluble in HCl is compared to 100 g of powdered

plant.

This test is used to distinguish between soluble physiological material and insoluble

substances like sand and earth.

193

A.1.1.18 Thin layer chromatography

Migration or solvent systems used are normally described for each plant.

The reagents used to treat the plates were: Dragendorffs reagent, AlCl3 and Godins

reagent.

Some results are as follows:

Dragendorffs reagent is used to indicate the presence of alkaloids.

AlCl3 is used to indicate the presence of flavonoids.

Godins reagent is used to indicate the presence of sterols, triterpenes, saponines, tannins,

cojmarins and chlorophyll.

A.1.1.19 Antioxidant activity

3 µg of each extract was placed on a thin layer chromatography plate. Migration systems

are described for each plant. The plates were treated with a solution of 2 mg/ml methanol

with DPPH.

Extracts with antioxidant activity will appear as white to yellow spots on a violet

background.

A.1.2 Reagents

Baljet’s reagent Picric acid 1 g

Ethanol 50º qsp 100 ml

Dragendorff’s reagent

Bismuth nitrate (powder) 20,8 g

Anhydrous sodium iodide 200 g

Iodine 38,10 g

Distilled water 600 ml

Stir for 30 minutes.

DPPH

1-1 difenyl 2 picril hydrazyl 2 mg/ml metanol

Fehling’s reagent

Solution A: CuSO4 35g


H2SO4 5 ml

194

Cool down and complete to 1 liter with distilled water.

Solution B: Seignettes salt 150 g


Cool down and add 300 ml non-carbonate washing powder. Complete to 1 liter with

distilled water.

Mix equal volumes of the the two solutions before use.

Godin’s reagent

Solution A:Vanilline 1 g

Ethanol 95 % 1000 ml

Solution B: HClO4 3 ml


Mix the two solutions before use.

The thin layer plates are first sprayed with the reagent, then with a 10% solution of

H2SO4. Coloured compounds appear after heating the plat up to 100 ºC.

Guignard’s reagent (preparation of ”picrosode” paper)

Picric acid 1 g

Sodium carbonate 10 g


Kedde’s reagent

3,5- dinitro benzoic acid 1 g


Raymond Marthoud’s reagent

1-3 M dinitrobenzene 1 g


Resorcine relevation solution

Resorcine 100 mg

Ethanol 50 ml

Phosphoric acid 50 ml

Valser-Mayer’s reagent

Potassium iodate 25 g

Mercuric chloride (HgCl) 6,77 g


195

Plant family Plant Author Year

Aizoaceae Glinus oppostifolius Bah 1998

Aizoaceae Limeum pterocarpum Bah 1998

Anacardiaceae Lannea microcarpa Sanogo 1999

Anacardiaceae Lannea velutina Field work 2004

Anacardiaceae Lannea velutina Field work 2004

Anacardiaceae Lannea velutina Kanta 1999

Anacardiaceae Lannea velutina Oulogeum 1999

Anacardiaceae Lannea velutina Samake 2000

Anacardiaceae Mangifera indica Aouissa 2002

Anacardiaceae Spondias mombin Sangare, D. 2003

Annonaceae Annona senegalensis Kanta 1999

Annonaceae Annona senegalensis Samake 2000

Araliaceae Cussonia barteri Bouare 2004

Araliaceae Cussonia barteri Oulogeum 1999

Arecaceae Borassus flabellifer Field work 2004

Asclepiadaceae Calotropis procera Field work 2004

Asclepiadaceae Leptadenia pyrotechnica Bah 1998

Asclpiadaceae Leptadenia hastata Field work 2004

Asteraceae Vernonia kotschyana Ba 1998

Asteraceae Vernonia kotschyana Bah 1998

Asteraceae Vernonia kotschyana Oulogeum 1999

Asteraceae Vernonia colorata Fane 2002

Balanitaceae Balanites aegyptiaca Ahamet 2003

Bignoniaceae Stereospermum kunthianum Kanta 1999

Bignoniaceae Stereospermum kunthianum Samake 2000

Bombacaceae Bombax costatum Sanogo 1999

Bombacaceae Bombax costatum Field work 2004

Caesalpiniaceae Afzelia africana Field work 2004

Caesalpiniaceae Burkea africana Kanta 1999

Caesalpiniaceae Burkea africana Samake 2000

Caesalpiniaceae Cassia nigricans Kanta 1999

Caesalpiniaceae Cassia nigricans Samake 2000

Caesalpiniaceae Cassia sieberiana Ekoumou 2004

Caesalpiniaceae Cassia sieberiana Fane 2002

Caesalpiniaceae Cassia sieberiana rp. Ekoumou 2004

Caesalpiniaceae Cordyla pinnata Sanogo 1999

Caesalpiniaceae Daniellia oliveri # 1 Sanogo 1999

Caesalpiniaceae Daniellia oliveri # 2 Sanogo 1999

Caesalpiniaceae Piliostigma thonningii Ekoumou 2004

Caesalpiniaceae Piliostigma thonningii rp. Ekoumou 2004

196

Capparidaceae Boscia senegalensis Ba 1998

Capparidaceae Boscia senegalensis Bah 1998

Capparidaceae Maerua crassifolia Bah 1998

Celastraceae Maytenus senegalensis Kanta 1999

Cochlospermaceae Cochlospermum tinctorium Kanta 1999

Cochlospermaceae Cochlospermum tinctorium Samake 2000

Combretaceae Combretum ghasalense # 1 Sanogo 1999

Combretaceae Combretum ghasalense # 2 Sanogo 1999

Combretaceae Combretum glutinosum Field work 2004

Combretaceae Combretum molle Field work 2004

Combretaceae Combretum molle Sanogo 1999

Combretaceae Pteleopsis suberosa Oulogeum 1999

Combretaceae Terminalia avicennoides Sanogo 1999

Combretaceae Terminalia macroptera Sanogo 1999

Convolvulaceae Ipomoea asarifola Field work 2004

Ebenaceae Diospyros abyssincia Oulogeum 1999

Euphorbiaceae Bridelia ferruginea Kanta 1999

Euphorbiaceae Euphorbia sudanica Field work 2004

Euphorbiaceae Ricinus communis Field work 2004

Euphorbiaceae Securinega virosa Sangare, D. 2003

Euphorbiaceae Securinega virosa Sangare, D. 2003

Fabaceae Afrormosia laxiflora Fane 2002

Fabaceae Afrormosia laxiflora Kanta 1999

Fabaceae Bauhinia thoningii Kanta 1999

Fabaceae Bauhinia thoningii Samake 2000

Fabaceae Ostryoderris stuhlmannii Kanta 1999

Fabaceae Ostryoderris stuhlmannii Samake 2000

Fabaceae Prosopis africana Ekoumou 2004

Fabaceae Prosopis africana rp. Ekoumou 2004

Fabaceae Pterocarpus erinaceus Field work 2004

Fabaceae Pterocarpus erinaceus Kanta 1999

Fabaceae Pterocarpus erinaceus Samake 2000

Fabaceae Pterocarpus lucens Field work 2004

Fabaceae Stylosanthes erecta Ekoumou 2004

Fabaceae Stylosanthes erecta (SEH) Ekoumou 2004

Fabaceae Stylosanthes erecta (SET) Ekoumou 2004

Flacourtiaceae Flacourtia flabescens Oulogeum 1999

Hypericaceae Psorospermum guineense Oulogeum 1999

Liliaceae Aloe buteneri Field work 2004

Lycoperdeae Podaxon aegyptiacus Samake 2000

Lythraceae Lawsonia inermis Ba 1998

197

Lythraceae Lawsonia inermis Oulogeum 1999

Meliaceae Khaya senegalensis Sanogo 1999

Meliaceae Trichilia emetica Fane 2002

Meliaceae Trichilia emetica Kanta 1999

Meliaceae Trichilia emetica Samake 2000

Meliaceae Trichilia emetica Timbo 2003

Mimosaceae Acacia ataxacantha Field work 2004

Mimosaceae Acacia dudgeoni - G3 Sanogo 1999

Mimosaceae Acacia senegal - G1 Sanogo 1999

Mimosaceae Acacia seyal - G2 Sanogo 1999

Mimosaceae Entada africana Ba 1998

Mimosaceae Entada africana Bah 1998

Mimosaceae Entada africana Oulogeum 1999

Mimosaceae Parkia biglobosa Field work 2004

Moraceae Ficus iteophila Kanta 1999

Moringaceae Moringa oleifera Chetima 2004

Olacaceae Ximenia americana Field work 2004

Olacaceae Ximenia americana Kanta 1999

Opiliaceae Opilia celtidifolia Field work 2004

Opiliaceae Opilia celtidifolia Sangare, D. 2003

Opiliaceae Opilia celtidifolia Sangare, D. 2003

Oxalidaceae Biophytum petersianum Field work 2004

Oxalidaceae Biophytum petersianum Kanta 1999

Oxalidaceae Biophytum petersianum Samake 2000

Papaveraceae Argemone mexicana Sangare, D. 2003

Papilionaceae Swartzia madagascariensis Fane 2002

Poaceae Zea mays Field work 2004

Polygalaceae Securidaca longepedunculata Tolo 2002

Rhamnaceae Ziziphus mauritania Oulogeum 1999

Rhamnaceae Ziziphus mauritania Yansambou 2002

Rhamnaceae Zizyhpus mucronata Ba 1998

Rubiaceae Canthium acutiflorum Sangare, D. 2003

Rubiaceae Canthium acutiflorum Sangare, D. 2003

Rubiaceae Crossopteryx febrifuga Kanta 1999

Rubiaceae Crossopteryx febrifuga Samake 2000

Rubiaceae Feretia apodanthera Sangare, D. 2003

Rubiaceae Feretia apodanthera Sangare, D. 2003

Rubiaceae Mitracarpus scaber Field work 2004

Rubiaceae Mitragyna inermis Bah 1998

Salvadoraceae Salvadora persica Bah 1998

Sapotaceae Butyrospermum parkii Field work 2004

http://www.zimbabweflora.co.zw/speciesdata/papilionac/index.php

http://www.museums.org.za/bio/plants/rubiaceae/index.htm

http://www.museums.org.za/bio/plants/rubiaceae/index.htm

198

Sterculiaceae Cola cordifolia Field work 2004

Sterculiaceae Cola cordifolia Kanta 1999

Sterculiaceae Cola cordifolia Samake 2000

Ulmaceae Celtis integrifolia Bah 1998

Vitaceae Cissus quadrangualris Ba 1998

Vitaceae Cissus quadrangularis Oulogeum 1999

A literature survey of studies performed by master ... · performed by master students at Département de Médecine Traditionelle (DMT) ... Avdeling for farmakognosi Kjemisk seksjon

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